Triazole derivativesas p2y14 receptor antagonists

ABSTRACT

Described are compounds, which are antagonists of the P2Y 14  receptor, for example, a compound of formula (I) in which ring A, R 1 , R 2 , R 3 , and n are as described herein. Also provided are dendron conjugates comprising the compounds, and methods of using the compounds, including a method of treating a disorder, such as inflammation, diabetes, insulin resistance, hyperglycemia, a lipid disorder, obesity, a condition associated with metabolic syndrome, and asthma, and a method of antagonizing P2 14  receptor activity in a cell.

CROSS-REFERENCE TO A RELATED APPLICATION

This patent application claims the benefit of U.S. Provisional PatentApplication No. 62/233,162, filed Sep. 25, 2015, which is incorporatedby reference for all purposes.

BACKGROUND OF THE INVENTION

Extracellular uridine-5′-diphosphate 1 and uridine-5′-diphosphoglucose 2(FIG. 1) activate the P2Y₁₄ receptor (P2Y₁₄R), a G protein-coupledreceptor (GPCR) of the δ-branch of Family A, to modulate function inmodels of inflammation, diabetes, asthma and other diseases (Lazarowskiet al., Mol. Pharmacol. 2015, 88(1), 151-160; and Abbracchio et al.,Pharmacol. Rev. 2006, 58, 281-341). This receptor subtype is a member ofthe P2Y₁₂R-like subfamily of nucleotide receptors, which inhibits theproduction of cyclic AMP through Gi protein. The P2Y₁₄R promoteshypersensitivity in microglial cells (Kobayashi et al., Glia 2012, 60,1529-1539), the mobility of neutrophils (Sesma et al., Am. J.Physiol.—Cell Physiol. 2012, 303, C490-C498), the release of mediatorsfrom mast cells (Gao et al., Biochem. Pharmacol. 2010, 79, 873-879),inflammation in renal intercalated cells (Azroyan et al., PLoS ONE 2015,10(3), e0121419. doi:10.1371/joumal.pone.0121419) and mixed effects ininsulin function (Xu et al., J. Immunol. 2012, 189(4), 1992-1999; andMeister et al., J. Biol. Chem. 2014, 289, 23353-23366). Thus, approachesto novel antagonists of nucleotide signaling at the P2Y₁₄R would bedesirable for exploration as novel therapeutics for treating diseasesassociated with modulating P2Y₁₄R.

Only a limited set of P2Y₁₄R antagonists are currently known. Severalchemotypes based on naphthoic acid and pyrido[4,3-d]pyrimidine have beenreported to provide potent P2Y₁₄R antagonists, but displayed low oralbioavailability (International Patent Application WO 2009/070873;Gauthier et al., Bioorg. Med. Chem. Lett. 2011, 21, 2836-2839; Guay etal., Bioorg. Med. Chem. Lett. 2011, 21, 2832-2835; and Robichaud et al.,Bioorg. Med. Chem. Lett. 2011, 21, 4366-4368).

Despite these efforts, there remains an unmet need for novel antagonistswith improved potency, selectivity, and/or bioavailability for thetreatment of disorders that respond to modulating P2Y₁₄R.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a compound of formula (I):

in which ring A, R¹, R², R³, and n are as described herein. Alsoprovided is a compound of formula (II),

in which ring A′, R^(1′), R^(2′), R^(3′), and n′ are as describedherein. It has been discovered that a compound defined herein iseffective in antagonizing P2Y₁₄R activity. It is envisioned that acompound of formula (I) or (II) is desirable for therapeuticapplications because the compound inhibits P2Y₁₄R to modulate functionin models of inflammation, diabetes, insulin resistance, hyperglycemia,a lipid disorder, obesity, a condition associated with metabolicsyndrome, and asthma.

The invention also provides a dendron conjugate that comprises acompound of formula (I) or (II), which can have a structure of formula(III) or (IV), respectively:

in which ring A, R¹, R³, X¹, X², X³, X⁴, X⁵, n, ring A′, R^(1′), R^(3′),X^(1′), X^(2′), X^(3′), X^(4′), X^(5′), and n′ are as described herein.

The compounds of formula (I) and (II) simultaneously reduce themolecular weight and avoid the high lipophilicity of the naphthoic acidand pyrido[4,3-d]pyrimidine derivative 3 (FIG. 1) that contributes toits low solubility and difficulty of purification (Robichaud et al.,Bioorg. Med. Chem. Lett. 2011, 21(14), 4366-4368).

Thus, the invention further provides a method for treating inflammation,diabetes, insulin resistance, hyperglycemia, a lipid disorder, obesity,a condition associated with metabolic syndrome, or asthma in a mammal inneed thereof, which comprises administering a therapeutically effectiveamount of a compound of formula (I) or (II) or a pharmaceuticallyacceptable salt thereof to the mammal.

Also provided is a method of antagonizing P2Y₁₄R activity in a cellcomprising administering a compound of formula (I) or (II), a conjugatethereof, or a pharmaceutically acceptable salt thereof to a cell,whereby activity of P2Y₁₄R is antagonized.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIG. 1 shows prior art agonist and antagonist ligand probes of theP2Y₁₄R.

FIG. 2 is a chemical scheme of the synthesis of triazolyl derivatives offormula (I) (24a-n) as P2Y₁₄R antagonists in accordance with anembodiment of the invention. Reagents and conditions: a. CH₃OH, SOCl₂, 0to 23° C. (98%); b. 1) Boc₂O, (C₂H₅)₃N, DMAP, DCM; 2) K₂CO₃, MeOH,reflux (70%); c. PdCl₂(DPPF) (DPPF:1,1′-bis(diphenylphosphino)ferrocene), AcOK, DMF, 95° C. (74%); d. TFA,90° C. (97%); e. H₂, Rh/C, 100 psi (98%); f. Pd(Ph₃P)₄, K₂CO₃, DME, 85°C. (71%); g. (CF₃CO)₂O, NEt₃, Et₂O; h. TFA, DCM (70%); i. 1) Ts-OH,NaNO₂, H₂O/ACN; 2) NaN₃, (83%); j. CuSO₄, sodium ascorbate (1M aq.) k.KOH (1M aq.).

FIG. 3 is a chemical scheme of the synthesis of alkynyl derivative 44 asa P2Y₁₄R antagonist. Reagents and conditions: a. CH₃OH, SOCl₂, 0 to 23°C. (33%); b. 1-ethynyl-4-(trifluoromethyl)benzene, CuI, PdCl₂(PPh₃)₂,DMF, (C₂H₅)₃N, 0 to 23° C. (81%); c. (CF₃SO₂)₂O, (C₂H₅)₃N, CH₂Cl₂ (98%);d. tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine-1-carboxylate,Pd(PPh₃)₄, K₂CO₃, DMF (67%); e. LiOH (aqueous 0.5M), CH₃OH reflux, thenHCl (aqueous 1M), pH 1 (28%).

FIG. 4A is a chemical scheme of the synthesis of ethyl4-(4-(1-(4-(1-(6-aminohexyl)-1H-1,2,3-triazol-4-yl)butyl)piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoate(34) and4-(4-(1-(4-(1-(6-aminohexyl)-1H-1,2,3-triazol-4-yl)butyl)piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoicacid (35) in accordance with an embodiment of the invention. Reagentsand conditions: a. TsCl, NEt₃, DMAP, CH₂Cl₂, r.t. 15 h (88%); b. LiBr,DMF, r.t., 12 h (82%); c. HBr 48% sol., 80° C., 20 h (61%); d. NaN₃,H₂O, reflux, 12 h (80%); e. SOCl₂, EtOH, 0° C. to r.t. (78%); f. K₂CO₃,DMF (92%); g. CuSO₄ (15 mol %), sodium ascorbate (45 mol %), t-BuOH:H₂O: CH₂Cl₂ (51%); h. LiOH (aqueous 0.5M), CH₃OH reflux, then HCl(aqueous 1M), pH 1 (21%).

FIG. 4B is an improved chemical scheme of the synthesis of prior artcompound6-amino-9-(2-carboxy-4-((6-(4-(4-(4-(4-(3-carboxy-6-(4-(trifluoromethyl)phenyl)-naphthalen-1-yl)phenyl)piperidin-1-yl)butyl)-1H-1,2,3-triazol-1-yl)hexyl)carbamoyl)-phenyl)-3-iminio-5-sulfo-3H-xanthene-4-sulfonate(4). Reagents and conditions: a. 1)N,N,N′,N′-tetramethyl-O—(N-succinimidyl)-uronium tetrafluoroborate(TSTU), N,N-diisopropylethylamine (DIPEA), dimethylformamide (DMF); 2)LiOH, 0.5 M, MeOH: H₂O; b. TSTU, DIPEA, DMF, water, 0° C.

FIG. 5 is a chemical scheme of the proposed synthesis of dendronprecursor 103d. X is O, NH, or CH₂, and n ranges from 0-36.

FIG. 6 is a chemical scheme of the proposed synthesis of dendronprecursors 107b and 107q. X is O, NH, or CH₂, and n ranges from 0-36.

FIG. 7 is a chemical scheme of the proposed synthesis of dendronprecursor 109.

FIG. 8 is a chemical scheme of the proposed synthesis of dendronprecursor 111.

FIGS. 9A-9C represent a single chemical scheme of proposed Strategy 1for preparing a dendron conjugate of formula (II).

FIGS. 10A-10C represent a single chemical scheme of proposed Strategy 2for preparing a dendron conjugate of formula (II).

FIG. 11 illustrates a dendron conjugate of formula (II) covalentlylinked to a quantum dot.

FIG. 12 illustrates a dendron conjugate of formula (II) covalentlylinked to a gold particle.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound of formula (I):

wherein

ring A is aryl, heteroaryl, or cycloalkyl;

R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

R² is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl,C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl, cyanoalkyl, aryl,heteroaryl, heterocycloalkyl, —(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or—(CH₂)_(m)heterocycloalkyl;

each R³ is the same or different and each is C₁-C₈ alkyl, C₂-C₈ alkenyl,C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴, —(CH₂)_(m)aryl,—(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl;

R⁴, R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl;and

m and n are the same or different and each is 0 or an integer from 1-5;

or a pharmaceutically acceptable salt thereof.

In certain compounds, ring A is phenyl, furanyl, thiazolyl, thienyl,pyrazolyl, pyridazinyl, pyridinyl, pyrazinyl, benzofuranyl, cyclopropyl,or cyclohexyl. In a preferred embodiment, ring A is phenyl.

In some embodiments, R¹ is —CO₂H. In other embodiments, R¹ is anysuitable bioisostere of carboxylate, particularly a bioisostere thatimproves at least one property of the compound of formula (I), such asimproves bioavailability and/or increases the binding affinity of acompound of formula (I) to P2Y₁₄R. Examples of a suitable bioisostere ofcarboxylate include

In any of the foregoing embodiments, R² is H or C₂-C₈ alkynyl.

In any of the foregoing embodiments, R³ is C₁-C₈ alkyl, hydroxy,hydroxyalkyl, C₁-C₈ alkoxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy,—CN, —NH₂, or —CO₂R⁴.

In any of the foregoing embodiments, n is 0 or 1 or 2.

In some embodiments of the compound of formula (I),

is selected from the group consisting of (optionally in combination withR¹ being —CO₂H and R² being H):

In another aspect of the invention is a compound of formula (II):

wherein

ring A′ is aryl, heteroaryl, or cycloalkyl;

R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl,cyanoalkyl, aryl, heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;

each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR^(5′)R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR^(5′)R^(6′),—NR^(5′)C(O)R⁴, —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl;

R^(4′), R^(5′), and R^(6′) are the same or different and each is H orC₁-C₈ alkyl; and

m′ and n′ are the same or different and each is 0 or an integer from1-5;

or a pharmaceutically acceptable salt thereof.

In certain compounds of formula (II), ring A′ is phenyl, furanyl,thiazolyl, thienyl, pyrazolyl, pyridazinyl, pyridinyl, pyrazinyl,benzofuranyl, cyclopropyl, or cyclohexyl. In a preferred embodiment,ring A′ is phenyl.

In some embodiments of formula (II), R^(1′) is —CO₂H. In otherembodiments, R^(1′) is any suitable bioisostere of carboxylate,particularly a bioisostere that improves at least one property of thecompound of formula (II), such as improves bioavailability and/orincreases the binding affinity of a compound of formula (II) to P2Y₁₄R.Examples of a suitable bioisostere of carboxylate include

In any of the foregoing embodiments, R^(2′) is H.

In any of the foregoing embodiments, R^(3′) is C₁-C₈ alkyl, hydroxy,hydroxyalkyl, C₁-C₈ alkoxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy,—CN, —NH₂, or —CO₂R^(4′).

In any of the foregoing embodiments, n′ is 0 or 1 or 2.

In any of the embodiments above, the term “alkyl” implies astraight-chain or branched alkyl substituent containing from, forexample, from about 1 to about 8 carbon atoms, e.g., from about 1 toabout 6 carbon atoms. Examples of alkyl group include methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl,isopentyl, n-hexyl, and the like.

The above definition of “alkyl” also applies wherever “alkyl” occurs aspart of a group, such as, e.g., in C₃-C₆ cycloalkylalkyl, hydroxyalkyl,haloalkyl (e.g., monohaloalkyl, dihaloalkyl, and trihaloalkyl),cyanoalkyl, aminoalkyl, alkylamino, dialkylamino, arylalkyl, etc. Thealkyl can be substituted or unsubstituted, as described herein. Even ininstances in which the alkyl is an alkylene chain (e.g., —(CH₂)_(n)—),the alkyl group can be substituted or unsubstituted.

In any of the embodiments above, the term “alkenyl,” as used herein,means a linear alkenyl substituent containing from, for example, about 2to about 8 carbon atoms (branched alkenyls are about 3 to about 8carbons atoms), e.g., from about 3 to about 6 carbon atoms (branchedalkenyls are about 3 to about 6 carbons atoms). In accordance with anembodiment, the alkenyl group is a C₂-C₄ alkenyl. Examples of alkenylgroup include ethenyl, allyl, 2-propenyl, 1-butenyl, 2-butenyl,1-pentenyl, 2-pentenyl, 3-pentenyl, 1-hexenyl, and the like. The alkenylcan be substituted or unsubstituted, as described herein.

In any of the embodiments above, the term “alkynyl,” as used herein,means a linear alkynyl substituent containing at least one carbon-carbontriple bond and from, for example, about 2 to about 8 carbon atoms(branched alkynyls are about 4 to about 12 carbons atoms), e.g., fromabout 2 to about 6 carbon atoms (branched alkynyls can be from about 4to about 8 carbon atoms), e.g., from about 2 to about 4 carbon atoms.Examples of such substituents include propynyl, propargyl, n-butynyl,pentynyl, isopentynyl, hexynyl, octynyl, and the like. The alkynyl canbe substituted or unsubstituted, as described herein.

In any of the embodiments above, the term “cycloalkyl,” as used herein,means a cyclic alkyl moiety containing from, for example, 3 to 6 carbonatoms or from 5 to 6 carbon atoms. Examples of such moieties includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. Thecycloalkyl can be substituted or unsubstituted, as described herein.

In any of the embodiments above, the term “hydroxy” refers to the group—OH.

In any of the embodiments above, the terms “alkoxy” and “cycloalkyloxy”embrace linear or branched alkyl and cycloalkyl groups, respectively,that are attached to a divalent oxygen. The alkyl and cycloalkyl groupsare the same as described herein. The term “aryloxy” refers tosubstituents that have an aryl group attached to divalent oxygen. Thearyl group is the same as described herein.

In any of the embodiments above, the term “halo” refers to a halogenselected from fluorine, chlorine, bromine, and iodine.

In any of the embodiments above, the term “aryl” refers to a mono-, bi-,or tricyclic carbocyclic ring system having one, two, or three aromaticrings, for example, phenyl, naphthyl, anthracenyl, or biphenyl. The term“aryl” refers to an unsubstituted or substituted aromatic carbocyclicmoiety, as commonly understood in the art, and includes monocyclic andpolycyclic aromatics such as, for example, phenyl, biphenyl, naphthyl,anthracenyl, pyrenyl, and the like. An aryl moiety generally containsfrom, for example, 6 to 30 carbon atoms, from 6 to 18 carbon atoms, from6 to 14 carbon atoms, or from 6 to 10 carbon atoms. It is understoodthat the term aryl includes carbocyclic moieties that are planar andcomprise 4n+2 π electrons, according to Hückel's Rule, wherein n=1, 2,or 3. The aryl can be substituted or unsubstituted, as described herein.

In any of the embodiments above, the term “heteroaryl” refers toaromatic 5 or 6 membered monocyclic groups, 9 or 10 membered bicyclicgroups, and 11 to 14 membered tricyclic groups which have at least oneheteroatom (O, S, or N) in at least one of the rings. Each ring of theheteroaryl group containing a heteroatom can contain one or two oxygenor sulfur atoms and/or from one to four nitrogen atoms provided that thetotal number of heteroatoms in each ring is four or less and each ringhas at least one carbon atom. The fused rings completing the bicyclicand tricyclic groups may contain only carbon atoms and may be saturated,partially saturated, or unsaturated. The nitrogen and sulfur atoms canoptionally be oxidized, and the nitrogen atoms can optionally bequaternized. Heteroaryl groups which are bicyclic or tricyclic mustinclude at least one fully aromatic ring but the other fused ring orrings may be aromatic or non-aromatic. The heteroaryl group can beattached at any available nitrogen or carbon atom of any ring.Illustrative examples of heteroaryl groups are pyridinyl, pyridazinyl,pyrimidyl, pyrazinyl, benzimidazolyl, triazinyl, imidazolyl, (1,2,3)-and (1,2,4)-triazolyl, pyrazinyl, tetrazolyl, furanyl, benzofuranyl,pyrrolyl, thienyl, isothiazolyl, thiazolyl, isoxazolyl, and oxadiazolyl.The heteroaryl can be substituted or unsubstituted, as described herein.

The term “heterocycloalkyl” means a stable, saturated, or partiallyunsaturated monocyclic, bicyclic, and spiro ring system containing 3 to7 ring members of carbon atoms and other atoms selected from nitrogen,sulfur, and/or oxygen. In an aspect, a heterocycloalkyl is a 5, 6, or7-membered monocyclic ring and contains one, two, or three heteroatomsselected from nitrogen, oxygen, and sulfur. The heterocycloalkyl can beattached to the parent structure through a carbon atom or through anyheteroatom of the heterocycloalkyl that results in a stable structure.Examples of such heterocycloalkyl rings are isoxazolyl, thiazolinyl,imidazolidinyl, piperazinyl, homopiperazinyl, pyrrolyl, pyrrolinyl,pyrazolyl, pyranyl, piperidyl, oxazolyl, and morpholinyl. Theheterocycloalkyl can be substituted or unsubstituted, as describedherein.

In other aspects, any substituent that is not hydrogen (e.g., C₁-C₈alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocycloalkyl, or heterocycloalkylalkyl) can be an optionallysubstituted moiety. The substituted moiety typically comprises at leastone substituent (e.g., 1, 2, 3, 4, 5, 6, etc.) in any suitable position(e.g., 1-, 2-, 3-, 4-, 5-, or 6-position, etc.). When an aryl group issubstituted with a substituent, e.g., halo, amino, alkyl, OH, alkoxy,and others, the aromatic ring hydrogen is replaced with the substituentand this can take place in any of the available hydrogens, e.g., 2, 3,4, 5, and/or 6-position wherein the 1-position is the point ofattachment of the aryl group in the compound of the present invention.Suitable substituents include, e.g., halo, alkyl, alkenyl, alkynyl,hydroxy, nitro, cyano, amino, alkylamino, alkoxy, aryloxy, aralkoxy,carboxyl, carboxyalkyl, carboxyalkyloxy, amido, alkylamido,haloalkylamido, aryl, heteroaryl, and heterocycloalkyl, as describedherein.

In any of the embodiments above, whenever a range of the number of atomsin a structure is indicated (e.g., a C₁₋₈, C₁₋₆, or C₁₋₄ alkyl,cycloalkyl, etc.), it is specifically contemplated that any sub-range orindividual number of carbon atoms falling within the indicated rangealso can be used. Thus, for instance, the recitation of a range of 1-8carbon atoms (e.g., C₁-C₈), 1-6 carbon atoms (e.g., C₁-C₆), 1-4 carbonatoms (e.g., C₁-C₄), 1-3 carbon atoms (e.g., C₁-C₃), or 2-8 carbon atoms(e.g., C₂-C₈) as used with respect to any chemical group (e.g., alkyl,cycloalkyl, etc.) referenced herein encompasses and specificallydescribes 1, 2, 3, 4, 5, 6, 7, and/or 8 carbon atoms, as appropriate, aswell as any sub-range thereof (e.g., 1-2 carbon atoms, 1-3 carbon atoms,1-4 carbon atoms, 1-5 carbon atoms, 1-6 carbon atoms, 1-7 carbon atoms,1-8 carbon atoms, 2-3 carbon atoms, 2-4 carbon atoms, 2-5 carbon atoms,2-6 carbon atoms, 2-7 carbon atoms, 2-8 carbon atoms, 3-4 carbon atoms,3-5 carbon atoms, 3-6 carbon atoms, 3-7 carbon atoms, 3-8 carbon atoms,4-5 carbon atoms, 4-6 carbon atoms, 4-7 carbon atoms, 4-8 carbon atoms,etc., as appropriate).

The subscript “n” represents the number of substituents, e.g., R³, inwhich each substituent, e.g., R³, can be the same or different. Thesubscripts “m,” “o,” and “q” represent the number of methylene repeatunits. The subscript “p” represents the number of repeat units of alinker unit. The subscripts m, n, and q can be the same or different andeach is either 0 or an integer from 1-5 (i.e., 1, 2, 3, 4, or 5). Thesubscript o is an integer from 1-5 (i.e., 1, 2, 3, 4, or 5). Thesubscript p is either 0 or an integer from 1-36 (i.e., 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36). When m, n, p, or q is 0,then the corresponding moiety, i.e., methylene group, repeat unit, orR³, is not present in the compound of formula (I) or (II).

In any of the embodiments above, the phrase “salt” or “pharmaceuticallyacceptable salt” is intended to include nontoxic salts synthesized fromthe parent compound which contains a basic or acidic moiety byconventional chemical methods. Generally, such salts can be prepared byreacting the free acid or base forms of these compounds with astoichiometric amount of the appropriate base or acid in water or in anorganic solvent, or in a mixture of the two. For example, an inorganicacid (e.g., hydrochloric acid, sulfuric acid, phosphoric acid, orhydrobromic acid), an organic acid (e.g., oxalic acid, malonic acid,citric acid, fumaric acid, lactic acid, malic acid, succinic acid,tartaric acid, acetic acid, trifluoroacetic acid, gluconic acid,ascorbic acid, methylsulfonic acid, or benzylsulfonic acid), aninorganic base (e.g., sodium hydroxide, potassium hydroxide, calciumhydroxide, magnesium hydroxide, or ammonium hydroxide), an organicbase(e.g., methylamine, diethylamine, triethylamine, triethanolamine,ethylenediamine, tris(hydroxymethyl)methylamine, guanidine, choline, orcinchonine), or an amino acid (e.g., lysine, arginine, or alanine) canbe used. Generally, nonaqueous media such as ether, ethyl acetate,ethanol, isopropanol, or acetonitrile are typical. Lists of suitablesalts are found in Remington's Pharmaceutical Sciences, 18th ed., MackPublishing Company, Easton, Pa., 1990, p. 1445, and Journal ofPharmaceutical Science, 66, 2-19 (1977). For example, they can be a saltof an alkali metal (e.g., sodium or potassium), alkaline earth metal(e.g., calcium), or ammonium of salt.

A compound of formula (I) or (II) can be prepared by any suitablemethod, including the methods described herein. For example, a compoundof formula (I) can be prepared by coupling a dioxaborolane derivative of3-amino-5-bromobenzoic acid with 4-(4-bromophenyl)piperidine. A specificexample of this method is set forth in FIG. 2, in which triazolylderivatives 24a-n were synthesized starting from 3-amino-5-bromobenzoicacid 12 and 4-(4-bromophenyl)piperidin-4-ol 16. The carboxylic group of12 was first converted to the methyl ester 13, and then the aminefunction was protected to give Boc-derivative 14a. A di-Boc side product14b accompanied the product that was initially isolated, and thisimpurity reverted to the desired mono-Boc product 14a upon heating themixture in refluxing MeOH in the presence of dilute K₂CO₃. Thepalladium-catalyzed condensation of arylbromide 14a withbis(pinacolato)-diboron under basic conditions afforded dioxaborolane15. The acid-catalyzed dehydration of 16 yielded derivative 17, whichwas reduced to provide compound 18. Derivative 19 was obtained bycoupling 18 with compound 15 under Suzuki conditions (Suzuki et al.,Angew. Chem. Int. Ed. 2011, 50 (30), 6722-6737). The conversion of theamino group of 19 to a trifluoroacetamide derivative 20 was accomplishedby using trifluoroacetic anhydride in the presence of base. Afterremoving the N-Boc protecting group of 20 to give compound 21, arylazide 22 was formed from an arenediazonium tosylate that was generatedin situ and subsequent addition of sodium azide (Kutonova et al.,Synthesis 2013, 45 (19), 2706-2710). In particular, the 1,2,3-triazolylderivative 23b was synthesized via a click reaction involving aryl azide22, 4-(trifluoromethyl)phenylacetylene 6b, Cu(II) salt and sodiumascorbate (Himo et al., J. Am. Chem. Soc. 2005, 127 (1), 210-216),followed by the one-pot hydrolysis of the trifluoroacetamide and theester in the presence of potassium hydroxide to yield 24b.

The present invention further provides a dendron conjugate, in which atleast one compound of formula (I) or (II) is linked to a dendron throughthe nitrogen atom on the piperidinyl group instead of R² or R^(2′). Insome instances, the dendron is attached to one compound of formula (I)or (II), whereas in other instances, the dendron is attached to morethan one compound of formula (I) or (II). The dendron conjugate canoptionally be attached to a particle as described herein.

In some embodiments, the dendron conjugate has a structure of formula(III)

or a pharmaceutically acceptable salt thereof, wherein

ring A is aryl, heteroaryl, or cycloalkyl;

R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or an isostere of carboxylate;

each R³ is the same or different and each is C₁-C₈ alkyl, C₂-C₈ alkenyl,C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴, —(CH₂)_(m)aryl,—(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl;

R⁴, R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl;

X¹ is selected from the group consisting of —(CH₂)_(o)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—;

X² is selected from the group consisting of

R⁷ is CH₂, NH, or O;

X³ is a dendron;

X⁴ is selected from the group consisting of —(CH₂)_(o)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X⁵ is a reactive sulfur-containing moiety;

m, n, and q are the same or different and each is 0 or an integer from1-5;

o is an integer from 1-5; and

p is 0 or an integer from 1-36;

wherein X⁵ is optionally linked to a particle.

In other embodiments, the dendron conjugate has a structure of formula(IV)

or a pharmaceutically acceptable salt thereof, wherein

ring A′ is aryl, heteroaryl, or cycloalkyl;

R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl,cyanoalkyl, aryl, heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;

each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR⁵R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR^(5′)R^(6′),—NR^(5′)C(O)R^(4′), —(CH₂)_(m)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl;

R^(4′), R^(5′), and R^(6′) are the same or different and each is H orC₁-C₈ alkyl;

X^(1′) is selected from the group consisting of —(CH₂)_(o′)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—;

X^(2′) is selected from the group consisting of

R⁷ is CH₂, NH, or O;

X^(3′) is a dendron;

X^(4′) is selected from the group consisting of —(CH₂)_(o′)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and;

X^(5′) is a reactive sulfur-containing moiety;

m′, n′, and q′ are the same or different and each is 0 or an integerfrom 1-5;

o′ is an integer from 1-5; and

p′ is 0 or an integer from 1-36;

wherein X^(5′) is optionally linked to a particle.

In certain dendron conjugates of formula (III) or (IV), ring A or ringA′ is phenyl, furanyl, thiazolyl, thienyl, pyrazolyl, pyridazinyl,pyridinyl, pyrazinyl, benzofuranyl, cyclopropyl, or cyclohexyl. In apreferred embodiment, ring A or ring A′ is phenyl.

In some embodiments of formula (III) or (IV), R¹ or R^(1′) is —CO₂H. Inother embodiments, R¹ or R^(1′) is any suitable bioisostere ofcarboxylate, particularly a bioisostere that improves at least oneproperty of conjugate, such as improves bioavailability and/or increasesthe binding affinity of the conjugate to P2Y₁₄R. Examples of a suitablebioisostere of carboxylate include

In any of the foregoing embodiments of formula (III) or (IV), R³ orR^(3′) is C₁-C₈ alkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, halo, C₁-C₈haloalkyl, C₁-C₈ haloalkoxy, —CN, —NH₂, or —CO₂R⁴.

In any of the foregoing embodiments of formula (III) or (IV), n or n′ is0 or 1 or 2.

In some embodiments of the compound of formula (III) or (IV), R¹ orR^(1′) is —CO₂H; and

is selected from the group consisting of

Dendrimers are classified as polymers; however, they are made frombranched monomers through the iterative organic synthesis by adding onelayer (i.e., generation) at each step to provide a symmetricalstructure. The solution conformation of higher generation dendrimers canclosely mimic the size and shape of a protein. Furthermore, dendrimersgenerally possess favorable characteristics: structural integrity,control of component functional groups—and their physical properties—bychemical synthesis, feasibility to conjugate multiple functional unitsat the peripheries and interiors, and a low enzymatic degradation rate.A dendron is similar to a dendrimer but without the symmetricalstructure due to a reactive functional group close to the core of thedendron. In general, the reactive functional group can be, for example,amino, hydroxyl, thiol, sulfone (e.g., —RSO₂R′), sulfinic acid (e.g.,—RSO(OH)), sulfonic acid (e.g., —RSO₂(OH)), thiocyanato, allyl,acetylenyl, carboxyl, halocarboxy (e.g., —OC(O)X), halo, formyl,haloformyl (e.g., —RC(O)X), carbonato, ester, alkoxy, amido (e.g.,—C(O)NRR′ or —NRC(O)R′), azido, azo, cyano, nitro, nitroso, or acombination thereof. In a preferred embodiment, the reactive functionalgroup on the dendron available for conjugation to the compound offormula (I) or (II) is an amino group. The dendron can be anionic orcationic.

The dendron can be of any suitable generation, e.g., from G2 to G10 ormore, including fractional generations, particularly G2 to G8, e.g., G2,G2.5, G3, G3.5, G4, G4.5, G5, G5.5, G6, G6.5, G7, or G7.5. For example,the half generations are carboxyl terminated and full generations areamine terminated. In preferred embodiments, the dendron is of G1, G2, orG3.

The conjugate of the invention can include any suitable dendron that canform a bond to the nitrogen on the piperidine ring of the compound offormula (I) or (II). In particular, the dendron can be apoly(amidoamine) (PAMAM) dendron, carboxyethylpolyamido (CEPAM) dendron,2,2-bis(hydroxyl-methyl)propionic acid (bis-MPA) dendron,poly(propyleneimine) (PPI) dendron, poly-L-lysine dendron,poly(etherhydroxylamine) (PEHAM) dendron, poly(esteramine) (PEA)dendron, polyglycerol dendron, and combinations thereof (e.g., PEHAM/PEAdendron). If desired, the dendron can be further functionalized toimprove at least one physicochemical property such as, e.g., watersolubility. For example, a biologically inactive pendant PEGylated chaincan be added to at least one suitable position of the dendron. Thependant PEGylated chain can have the formula

in which R⁸ is CH₂, NH, or O; R⁹ is —NH₂ or —CO₂H; and p is 0 or aninteger from 1-36.

In some preferred aspects, the dendron is a CEPAM dendron that isoptionally functionalized in at least one position to include the moiety

in which R⁸ is CH₂, NH, or O; R⁹ is —NH₂ or —CO₂H; and p is 0 or aninteger from 1-36. For example, the CEPAM dendrons can be formed byunits of 4-amino-4-(2-carboxyethyl)heptanedioic acid, with the followingstructures:

The conjugate is optionally linked to a particle through a sulfur atomof X⁵ or X^(5′). Typically, the particle is any nanoparticle ormicroparticle that has a surface suitable for bonding to an organicmoiety. For example, the particle can be a quantum dot, a non-metallicparticle, or a metallic particle. The quantum dot can comprise, forexample, a core of the formula MX, in which M is cadmium, zinc, mercury,aluminum, lead, tin, gallium, indium, thallium, magnesium, calcium,strontium, barium, copper, and mixtures or alloys thereof; and X issulfur, selenium, tellurium, nitrogen, phosphorus, arsenic, antimony ormixtures thereof. Suitable examples of the quantum dot include leadsulfide, lead selenide, lead telluride, zinc cadmium, zinc selenide,zinc sulfide, zinc telluride, cadmium selenide, cadmium sulfide, indiumarsenide, indium phosphide, thallium arsenide, thallium nitride,thallium phosphide, thallium antimonide, and alloys of the foregoing.The non-metallic particle can comprise, for example, silica, titania,alumina, germania, calcium carbonate, barium sulfate, or a combinationthereof. The metallic particle can comprise, for example, gold, silver,platinum, palladium, ruthenium, copper, iron oxide, gallium selenide,indium selenide, lead selenide, cadmium sulfide, lead sulfide, or acombination thereof. The size of the particle is not particularlyimportant but can range from 1 nm to 1,000 μm (e.g., 1 nm to 10 nm, 10nm to 100 nm, 50 nm to 100 nm, 100 nm to 1 μm, 1 μm to 50 μm, 50 μm to250 μm, 250 μm to 500 μm, 500 μm to 750 μm, 500 μm to 1,000 μm). Any twoof the foregoing endpoints can be used in combination to define aclose-ended range.

In order to bond to the particle, X⁵ or X^(5′) is an organic group thatincludes a reactive sulfur-containing moiety. X⁵ or X^(5′) can be, forexample, a bifunctional linker. Bifunctional linkers are known in theart (e.g., Sigma-Aldrich, St. Louis, Mo.). The bifunctional linkercomprises any moiety that can form a chemical bond between the particleand X⁴ or X^(4′). The linker can be of any suitable charge, lengthand/or rigidity, but preferably the bifunctional linker is derived froma compound comprising a sulfur-containing moiety (e.g., thio) and asecond reactive group, such as an amino group, hydroxyl, thio, a halogroup, a carboxyl group, aryl, heteroaryl, or heterocyclyl group, priorto reaction with the particle, X⁴ or X^(4′), and/or the dendron.Examples of a suitable heteroaryl include

Preferably, these groups are at the terminal ends of the bifunctionallinker. Examples of the linker include, e.g., 2-(boc-amino)ethanethiol,biphenyl-4,4′-dicarbodithioic acid, 4-mercapto-1-butanol,6-mercapto-1-hexanol, [1,1′-biphenyl]-4,4′-dithiol, cysteine, and lipoicacid. In a preferred embodiment, X⁵ or X^(5′) is a residue of lipoicacid.

The methods described herein comprise using a compound of formula (I) or(II) or a conjugate or a pharmaceutically acceptable salt thereof in theform of a pharmaceutical composition. In particular, a pharmaceuticalcomposition will comprise (i) at least one compound of formula (I) or(II) or a conjugate thereof, or a pharmaceutically acceptable saltthereof and (ii) a pharmaceutically acceptable carrier. Thepharmaceutically acceptable excipients described herein, for example,vehicles, adjuvants, carriers or diluents, are well-known to those whoare skilled in the art and are readily available to the public.Typically, the pharmaceutically acceptable carrier is one that ischemically inert to the active compounds and one that has no detrimentalside effects or toxicity under the conditions of use.

The pharmaceutical compositions can be administered as oral, sublingual,transdermal, subcutaneous, topical, absorption through epithelial ormucocutaneous linings, intravenous, intranasal, intraarterial,intramuscular, intratumoral, peritumoral, interperitoneal, intrathecal,rectal, vaginal, or aerosol formulations. In some aspects, thepharmaceutical composition is administered orally or intravenously.

In accordance with any of the embodiments, the compound of formula (I)or (II) or a pharmaceutically acceptable salt thereof can beadministered orally to a subject in need thereof. Formulations suitablefor oral administration can include (a) liquid solutions, such as aneffective amount of the compound dissolved in diluents, such as water,saline, or orange juice and include an additive, such as cyclodextrin(e.g., α-, β-, or γ-cyclodextrin, hydroxypropyl cyclodextrin) orpolyethylene glycol (e.g., PEG400); (b) capsules, sachets, tablets,lozenges, and troches, each containing a predetermined amount of theactive ingredient, as solids or granules; (c) powders; (d) suspensionsin an appropriate liquid; and (e) suitable emulsions and gels. Liquidformulations may include diluents, such as water and alcohols, forexample, ethanol, benzyl alcohol, and the polyethylene alcohols, eitherwith or without the addition of a pharmaceutically acceptablesurfactant, suspending agent, or emulsifying agent. Capsule forms can beof the ordinary hard- or soft-shelled gelatin type containing, forexample, surfactants, lubricants, and inert fillers, such as lactose,sucrose, calcium phosphate, and cornstarch. Tablet forms can include oneor more of lactose, sucrose, mannitol, corn starch, potato starch,alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum,colloidal silicon dioxide, croscarmellose sodium, talc, magnesiumstearate, calcium stearate, zinc stearate, stearic acid, and otherexcipients, colorants, diluents, buffering agents, disintegratingagents, moistening agents, preservatives, flavoring agents, andpharmacologically compatible carriers. Lozenge forms can comprise theactive ingredient in a flavor, usually sucrose and acacia or tragacanth,as well as pastilles comprising the active ingredient in an inert base,such as gelatin and glycerin, or sucrose and acacia, emulsions, gels,and the like containing, in addition to the active ingredient, suchcarriers as are known in the art.

Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.The compound of formula (I) or (II) or a salt thereof can beadministered in a physiologically acceptable diluent in a pharmaceuticalcarrier, such as a sterile liquid or mixture of liquids, includingwater, saline, aqueous dextrose and related sugar solutions, an alcohol,such as ethanol, isopropanol, or hexadecyl alcohol, glycols, such aspropylene glycol or polyethylene glycol, glycerol ketals, such as2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, such aspoly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester orglyceride, or an acetylated fatty acid glyceride with or without theaddition of a pharmaceutically acceptable surfactant, such as a soap ora detergent, suspending agent, such as pectin, carbomers,methylcellulose, hydroxypropylmethylcellulose, orcarboxymethylcellulose, or emulsifying agents and other pharmaceuticaladjuvants.

Oils, which can be used in parenteral formulations include petroleum,animal, vegetable, or synthetic oils. Specific examples of oils includepeanut, soybean, sesame, cottonseed, corn, olive, petrolatum, andmineral. Suitable fatty acids for use in parenteral formulations includeoleic acid, stearic acid, and isostearic acid. Ethyl oleate andisopropyl myristate are examples of suitable fatty acid esters. Suitablesoaps for use in parenteral formulations include fatty alkali metal,ammonium, and triethanolamine salts, and suitable detergents include (a)cationic detergents such as, for example, dimethyl dialkyl ammoniumhalides, and alkyl pyridinium halides, (b) anionic detergents such as,for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether,and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergentssuch as, for example, fatty amine oxides, fatty acid alkanolamides, andpolyoxyethylene-polypropylene copolymers, (d) amphoteric detergents suchas, for example, alkyl-beta-aminopropionates, and 2-alkyl-imidazolinequaternary ammonium salts, and (3) mixtures thereof.

The parenteral formulations will typically contain from about 0.5 toabout 25% by weight of the inhibitors in solution. Suitablepreservatives and buffers can be used in such formulations. In order tominimize or eliminate irritation at the site of injection, suchcompositions may contain one or more nonionic surfactants having ahydrophile-lipophile balance (HLB) of from about 12 to about 17. Thequantity of surfactant in such formulations ranges from about 5 to about15% by weight. Suitable surfactants include polyethylene sorbitan fattyacid esters, such as sorbitan monooleate and the high molecular weightadducts of ethylene oxide with a hydrophobic base, formed by thecondensation of propylene oxide with propylene glycol. The parenteralformulations can be presented in unit-dose or multi-dose sealedcontainers, such as ampoules and vials, and can be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example, water, for injections, immediatelyprior to use. Extemporaneous injection solutions and suspensions can beprepared from sterile powders, granules, and tablets of the kindpreviously described.

The inhibitors may be made into injectable formulations. Therequirements for effective pharmaceutical carriers for injectablecompositions are well known to those of ordinary skill in the art. SeePharmaceutics and Pharmacy Practice, J. B. Lippincott Co., Philadelphia,Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbookon Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986).

Topically applied compositions are generally in the form of liquids(e.g., mouthwash), creams, pastes, lotions and gels. Topicaladministration includes application to the oral mucosa, which includesthe oral cavity, oral epithelium, palate, gingival, and the nasalmucosa. In some embodiments, the composition contains at least oneactive component and a suitable vehicle or carrier. It may also containother components, such as an anti-irritant. The carrier can be a liquid,solid or semi-solid. In embodiments, the composition is an aqueoussolution, such as a mouthwash. Alternatively, the composition can be adispersion, emulsion, gel, lotion or cream vehicle for the variouscomponents. In one embodiment, the primary vehicle is water or abiocompatible solvent that is substantially neutral or that has beenrendered substantially neutral. The liquid vehicle can include othermaterials, such as buffers, alcohols, glycerin, and mineral oils withvarious emulsifiers or dispersing agents as known in the art to obtainthe desired pH, consistency and viscosity. It is possible that thecompositions can be produced as solids, such as powders or granules. Thesolids can be applied directly or dissolved in water or a biocompatiblesolvent prior to use to form a solution that is substantially neutral orthat has been rendered substantially neutral and that can then beapplied to the target site. In embodiments of the invention, the vehiclefor topical application to the skin can include water, bufferedsolutions, various alcohols, glycols such as glycerin, lipid materialssuch as fatty acids, mineral oils, phosphoglycerides, collagen, gelatinand silicone based materials.

The compound of formula (I) or (II) or a pharmaceutically acceptablesalt thereof, alone or in combination with other suitable components,can be made into aerosol formulations to be administered via inhalation.These aerosol formulations can be placed into pressurized acceptablepropellants, such as dichlorodifluoromethane, propane, nitrogen, and thelike. They also may be formulated as pharmaceuticals for non-pressuredpreparations, such as in a nebulizer or an atomizer.

The dose administered to the mammal, particularly a human and othermammals, in accordance with the present invention should be sufficientto affect the desired response. One skilled in the art will recognizethat dosage will depend upon a variety of factors, including the age,condition or disease state, predisposition to disease, genetic defect ordefects, and body weight of the mammal. The size of the dose will alsobe determined by the route, timing and frequency of administration aswell as the existence, nature, and extent of any adverse side-effectsthat might accompany the administration of a particular inhibitor andthe desired effect. It will be appreciated by a person of skill in theart that various conditions or disease states may require prolongedtreatment involving multiple administrations.

The inventive methods comprise administering an effective amount of acompound of formula (I) or (II) or a pharmaceutically acceptable saltthereof. An “effective amount” means an amount sufficient to show ameaningful benefit in an individual, e.g., reducing inflammation,reducing the risk of developing a particular condition described herein,delaying the onset of a particular condition described herein. Themeaningful benefit observed in the mammal can be to any suitable degree(10, 20, 30, 40, 50, 60, 70, 80, 90% or more). In some aspects, one ormore symptoms of the disease to be treated are prevented, reduced,halted, or eliminated subsequent to administration of a compound offormula (I) or (II) or a pharmaceutically acceptable salt thereof,thereby effectively treating the disease to at least some degree.

Effective amounts may vary depending upon the biological effect desiredin the individual, condition to be treated, and/or the specificcharacteristics of the compound of formula (I) or (II) or apharmaceutically acceptable salt thereof and the individual. In thisrespect, any suitable dose of the compound of formula (I) or (II) or apharmaceutically acceptable salt thereof can be administered to themammal (e.g., human), according to the type of disorder to be treated.Various general considerations taken into account in determining the“effective amount” are known to those of skill in the art and aredescribed, e.g., in Gilman et al., eds., Goodman And Gilman's: ThePharmacological Bases of Therapeutics, 8th ed., Pergamon Press, 1990;and Remington's Pharmaceutical Sciences, 17th Ed., Mack Publishing Co.,Easton, Pa., 1990, each of which is herein incorporated by reference.The dose of the compound of formula (I) or (II) or a pharmaceuticallyacceptable salt thereof desirably comprises about 0.1 mg per kilogram(kg) of the body weight of the mammal (mg/kg) to about 400 mg/kg (e.g.,about 0.75 mg/kg, about 5 mg/kg, about 30 mg/kg, about 75 mg/kg, about100 mg/kg, about 200 mg/kg, or about 300 mg/kg). In another embodiment,the dose of the compound of formula (I) or (II) or salt thereofcomprises about 0.5 mg/kg to about 300 mg/kg (e.g., about 0.75 mg/kg,about 5 mg/kg, about 50 mg/kg, about 100 mg/kg, or about 200 mg/kg),about 10 mg/kg to about 200 mg/kg (e.g., about 25 mg/kg, about 75 mg/kg,or about 150 mg/kg), or about 50 mg/kg to about 100 mg/kg (e.g., about60 mg/kg, about 70 mg/kg, or about 90 mg/kg). Any two of the foregoingendpoints can be used in combination to define a close-ended range.

In an aspect, a compound formula (I) or (II) antagonizes P2Y₁₄R activityin a cell. The method comprises administering a compound of formula (I)or (II) or a conjugate or a pharmaceutically acceptable salt thereof toa cell. The only requirement is that the cell expresses P2Y₁₄R and canbe from any suitable tissue (e.g., muscle tissue, nervous tissue,connective tissue, or epithelial tissue). The tissue can be from anyorgan, including the head, neck, eye, skin, mouth, throat, esophagus,chest, bone, lung, colon, sigmoid, rectum, stomach, prostate, breast,ovaries, kidney, liver, pancreas, brain, intestine (e.g.,gastrointestinal), heart, or adrenals. The antagonism of P2Y₁₄R can bemeasured by any method, including the assay described herein.

P2Y₁₄R has been implicated in immune and inflammatory responses (Barrettet al., Mol Pharmacol, 2013, 84, 41-49). Moreover, P2Y₁₄R has been foundto play a role in the initiation of inflammation and insulin resistancein obesity (Xu et al., J. Immunol. 2012, 189(4), 1992-1999).Accordingly, without wishing to be bound by any theory, it is believedthat antagonizing P2Y₁₄R can be a viable pathway to treating disordersassociated with inflammation. Accordingly, the present inventionprovides a method for treating a disorder responsive to antagonism ofP2Y₁₄R, such as inflammation, diabetes (e.g., type 2 diabetes), insulinresistance, hyperglycemia, a lipid disorder, obesity, a conditionassociated with metabolic syndrome, or asthma, in a mammal in needthereof which comprises administering a therapeutically effective amountof a compound of formula (I) or (II) or a pharmaceutically acceptablesalt thereof to the mammal. A condition associated with metabolicsyndrome includes, e.g., obesity, dyslipidemia, hyperglycermia,decreased high density lipoprotein (HDL), elevated triglycerides, andelevated blood pressure.

For purposes of the present invention, the term “mammal” typicallyincludes, but is not limited to, the order Rodentia, such as mice, andthe order Logomorpha, such as rabbits. In some aspects, the mammals arefrom the order Camivora, including Felines (cats) and Canines (dogs),Artiodactyla, including Bovines (cows) and Swines (pigs) or of the orderPerssodactyla, including Equines (horses). In some aspects, the mammalsare of the order Primates, Ceboids, or Simioids (monkeys) or of theorder Anthropoids (humans and apes). In embodiments of the invention,the mammal is a human.

The invention is further illustrated by the following embodiments.

(1) A compound of formula (I):

wherein

ring A is aryl, heteroaryl, or cycloalkyl;

R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or an isostere of carboxylate;

R² is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl,C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl, cyanoalkyl, aryl,heteroaryl, heterocycloalkyl, —(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or—(CH₂)_(m)heterocycloalkyl; each R³ is the same or different and each isC₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl,C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈haloalkoxy, —CN, —NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴,—(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl;

R⁴, R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl;

m and n are the same or different and each is 0 or an integer from 1-5;

or a pharmaceutically acceptable salt thereof.

(2) The compound of embodiment (1), wherein ring A is phenyl, furanyl,thiazolyl, thienyl, pyrazolyl, pyridazinyl, pyridinyl, pyrazinyl,benzofuranyl, cyclopropyl, or cyclohexyl, or a pharmaceuticallyacceptable salt thereof.

(3) The compound of embodiment (2), wherein ring A is phenyl, or apharmaceutically acceptable salt thereof.

(4) The compound of any one of embodiments (1)-(3), wherein R¹ is —CO₂H,or a pharmaceutically acceptable salt thereof.

(5) The compound of any one of embodiments (1)-(3), wherein R¹ is abioisostere of carboxylate selected from the group consisting of

or a pharmaceutically acceptable salt thereof.

(6) The compound of any one of embodiments (1)-(5), wherein R² is H orC₂-C₈ alkynyl, or a pharmaceutically acceptable salt thereof.

(7) The compound of any one of embodiments (1)-(6), wherein R³ is C₁-C₈alkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, halo, C₁-C₈ haloalkyl, C₁-C₈haloalkoxy, —CN, —NH₂, or —CO₂R⁴, or a pharmaceutically acceptable saltthereof.

(8) The compound of any one of embodiments (1)-(7), wherein n is 1 or 2.

(9) The compound of any one of embodiments (1)-(6), wherein n is 0.

(10) The compound of embodiment (1), wherein R¹ is —CO₂H; R² is H; and

is selected from the group consisting of

or a pharmaceutically acceptable salt thereof.

(11) A compound of formula (II):

wherein

ring A′ is aryl, heteroaryl, or cycloalkyl;

R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl,cyanoalkyl, aryl, heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;

each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR^(5′)R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR^(5′)R^(6′),—NR^(5′)C(O)R^(4′), —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl;

R^(4′), R^(5′), and R^(6′) are the same or different and each is H orC₁-C₈ alkyl; and

m′ and n′ are the same or different and each is 0 or an integer from1-5;

or a pharmaceutically acceptable salt thereof.

(12) A dendron conjugate comprising a compound of any one of embodiments(1)-(11), wherein the compound is linked to a dendron through thenitrogen atom on the piperidinyl group instead of R² or R^(2′).

(13) The dendron conjugate of embodiment (12), wherein the dendron isattached to more than one compound of formula (I) or formula (II).

(14) A conjugate of formula (III)

or a pharmaceutically acceptable salt thereof, wherein

ring A is aryl, heteroaryl, or cycloalkyl;

R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

each R³ is the same or different and each is C₁-C₈ alkyl, C₂-C₈ alkenyl,C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴, —(CH₂)_(m)aryl,—(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl;

R⁴, R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl;

X¹ is selected from the group consisting of —(CH₂)_(o)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—;

X² is selected from the group consisting of

R⁷ is CH₂, NH, or O;

X³ is a dendron;

X⁴ is selected from the group consisting of —(CH₂)_(o)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X⁵ is a reactive sulfur-containing moiety;

m, n, and q are the same or different and each is 0 or an integer from1-5;

o is an integer from 1-5; and

p is 0 or an integer from 1-36;

wherein X⁵ is optionally linked to a particle.

(15) The conjugate of embodiment (14), wherein the dendron iscarboxyethylpolyamido (CEPAM) dendron that is optionally functionalizedin at least one position to include the moiety

wherein R⁸ is CH₂, NH, or O, and R⁹ is —NH₂ or —CO₂H.

(16) The conjugate of embodiment (15), wherein the dendron is ofgeneration G1, G2, or G3.

(17) The conjugate of any one of embodiments (14)-(16) that is linked toa particle through a sulfur atom of X⁵.

(18) The conjugate of embodiment (17), wherein the particle is a quantumdot, a non-metallic particle, or a metallic particle.

(19) The conjugate of embodiment (18), wherein the metallic particlecomprises gold, silver, platinum, ruthenium, iron oxide, galliumselenide, indium selenide, lead selenide, cadmium sulfide, lead sulfide,or a combination thereof.

(20) The conjugate of any one of embodiments (14)-(19), wherein X⁵comprises a residue of lipoic acid.

(21) A dendron conjugate of formula (IV)

or a pharmaceutically acceptable salt thereof, wherein

ring A′ is aryl, heteroaryl, or cycloalkyl;

R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate;

R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl,cyanoalkyl, aryl, heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;

each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR^(5′)R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR^(5′)R^(6′),—NR^(5′)C(O)R^(4′), —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl;

R^(4′), R^(5′), and R^(6′) are the same or different and each is H orC₁-C₈ alkyl;

X^(1′) is selected from the group consisting of —(CH₂)_(o′)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—;

X^(2′) is selected from the group consisting of

R^(7′) is CH₂, NH, or O;

X^(3′) is a dendron;

X^(4′) is selected from the group consisting of —(CH₂)_(o′)—, —C(O)—,—C(O)NH—, —OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X^(5′) is a reactive sulfur-containing moiety;

m′, n′, and q′ are the same or different and each is 0 or an integerfrom 1-5;

o′ is an integer from 1-5; and

p′ is 0 or an integer from 1-36;

wherein X^(5′) is optionally linked to a particle.

(22) A pharmaceutical composition comprising (i) at least one compoundof any one of embodiments (1)-(11), a conjugate of any one ofembodiments (12)-(21), or a pharmaceutically acceptable salt thereof and(ii) a pharmaceutically acceptable carrier.

(23) A method of antagonizing P2Y₁₄ receptor (P2Y₁₄R) activity in a cellcomprising administering a compound of any one of embodiments (1)-(11),a conjugate of any one of embodiments (12)-(21), or a pharmaceuticallyacceptable salt thereof to a cell, whereby activity of P2Y₁₄R isantagonized.

(24) A method for treating inflammation, diabetes, insulin resistance,hyperglycemia, a lipid disorder, obesity, a condition associated withmetabolic syndrome, or asthma in a mammal in need thereof whichcomprises administering a therapeutically effective amount of a compoundof any one of embodiments (1)-(11) or a pharmaceutically acceptable saltthereof to the mammal.

The following examples further illustrate the invention but, of course,should not be construed as in any way limiting its scope.

EXAMPLES

The proton and carbon nuclear magnetic resonance spectra were recordedusing a Bruker 400 MHz NMR spectrometer. Purification of final compoundswas performed by preparative high performance liquid chromatography(HPLC) (Column: Luna 5 μm C18(2) 100 Å, LC Column 250×4.6 mm). Method AEluant: 0.1% trifluoroacetic acid (TFA) in water-CH₃CN from 100:0 to70:30 in 45 min with a flow rate of 5 mL/min. Method B: Method B Eluant:10 mM triethylammonium acetate (TEAA) buffer —CH₃CN from 80:20 to 20:80in 40 min, then 10 mM TEAA buffer —CH₃CN from 20:80 to 0:100 in 10 minwith a flow rate of 5 mL/min. Purities of all tested compounds were≥95%, as estimated by analytical HPLC (Column: ZORBAX™ Eclipse 5 μmXDB-C18 analytical column, 150×4.6 mm; Agilent Technologies, Inc., SantaClara, Calif.). Peaks were detected by UV absorption (254 nm) using adiode array detector. All derivatives tested for biological activityshowed >95% purity in the HPLC systems. Analytical thin-layerchromatography was carried out on SIGMA-ALDRICH™ thin layerchromatography (TLC) plates and compounds were visualized with UV lightat 254 nm. Silica gel flash chromatography was performed using 230-400mesh silica gel. Unless noted otherwise, reagents and solvents werepurchased from Sigma-Aldrich (St. Louis, Mo.). Low-resolution massspectrometry was performed with a JEOL SX102 spectrometer (Peabody,Mass.) with 6-kV Xe atoms following desorption from a glycerol matrix oron an Agilent LC/MS 1100 MSD (Santa Clara, Calif.), with a WatersATLANTIS™ C18 column (Milford, Mass.). High-resolution massspectroscopic (HRMS) measurements were performed on a proteomicsoptimized WATERS™ MICROMASS™ Q-TOF-2™ using external calibration withpolyalanine.

Example 1

This example demonstrates the synthesis of the intermediate methyl3-amino-5-bromobenzoate (13) in an embodiment of the invention (FIG. 2).

3-Bromo-5-aminobenzoic acid 12 (1.0 g, 4.62 mmol) was stirred inmethanol (15 mL) with ice cooling, and the yellow solution was treatedwith thionyl chloride (4.00 mL, 55.0 mmol) dropwise over 20 min. Theresulting mixture was allowed to warm up to room temperature and leftstirring overnight. The reaction mixture was quenched with aqueoussaturated NaHCO₃ solution at 0° C. The solvent was then removed undervacuum, and the residue was suspended in ethyl acetate (200 mL). Theorganic phase was washed with brine (100 mL), dried (Na₂SO₄) andconcentrated to afford the title compound as a yellow solid (1.08 g,98%). ¹H NMR (400 MHz, Methanol-d) δ 7.10 (t, J=1.6 Hz, 1H), 6.83 (t,J=1.6 Hz, 1H), 6.57 (t, J=1.6 Hz, 1H), 3.46 (s, 3H). 13C NMR (100 MHz,CDCl₃) δ: 52.3, 114.6, 121.6, 122.3, 122.9, 132.6, 147.7, 166.0. m/z(ESI, MH⁺) 231. ESI-HRMS (MH⁺) calcd. for C₈H₈BrNO₂ 229.9817, found229.9818. HPLC purity 98.8% (R_(t)=12.3 min).

Example 2

This example demonstrates the synthesis of the intermediate methyl3-bromo-5-((tert-butoxycarbonyl)amino)benzoate (14) in an embodiment ofthe invention (FIG. 2).

To a solution of 13 (3.73 g, 16.2 mmol) in DCM (40 mL), Boc₂O (4.2 g,19.4 mmol) and DMAP (1.9 g, 16.2 mmol) were sequentially added with icecooling bath. The resulting mixture was allowed to stir at 0° C. for 2h. The solvent was removed under vacuum, and the resulting residue waspurified by silica gel chromatography using as eluant hexane/EtOAc(75:25) to afford the title compound as a white solid (4.3 g, 80%). ¹HNMR (400 MHz, CDCl₃): δ 7.98 (s, 1H), 7.82 (s, 1H), 7.80 (s, 1H), 6.60(s, 1H), 3.91 (s, 3H), 1.52 (s, 9H). ¹³C NMR (100 MHz, MeOD): δ 165.6,153.3, 141.3, 132.1, 125.4, 124.7, 121.9, 117.5, 51.5, 27.2. MS (ESI,m/z) 331 [M+H]+; ESI-HRMS calcd. m/z for C₁₃H₁₆BrNO₄ 329.0263, found329.0260 [M+H]⁺. HPLC purity 99.6% (R_(t)=20.14 min).

Example 3

This example demonstrates the synthesis of the intermediate methyl3-((tert-butoxycarbonyl)amino)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate(15) in an embodiment of the invention (FIG. 2).

A solution of 14 (1.36 g, 4.11 mmol), bis(pinacolato)diboron (1.25 g,4.94 mmol), KOAc (1.21 g, 12.3 mmol) in dry DMF (12 mL) was degassedwith N₂ for 30 min. Then, PdCl₂(DPPF) (DPPF:1,1′-bis(diphenylphosphino)ferrocene) (0.30 g, 0.41 mmol) was addedwhile continuing degassing for additional 5 min. The reaction mixturewas heated at 95° C. and left stirring overnight. After cooling, theresulting mixture was suspended in EtOAc and filtered through CELITE™(Sigma Aldrich, St. Louis, Mo.). The solvent was removed under vacuumleaving a black residue, which was purified by silica gel chromatographyusing as eluant hexane:EtOAc (75:25). The title compound was obtained asa white solid (1.1 g, 74%). ¹H NMR (400 MHz, Chloroform-d) δ 8.16 (t,J=2.0 Hz, 1H), 8.14 (t, J=2.0 Hz, 1H), 7.90 (t, J=2.0 Hz, 1H), 6.53 (s,1H), 3.90 (s, 3H), 1.52 (s, 9H), 1.34 (s, 12H). 13C NMR (100 MHz, CDCl₃)δ: 24.9, 28.3, 52.1, 84.1, 122.2, 128.9, 130.0, 138.2, 152.6, 166.9. m/z(ESI, MH⁺) 378. ESI-HRMS (MH⁺) calcd. for C₁₉H₂₈BNO₆ 376.2046, found376.2049.

Example 4

This example demonstrates the synthesis of the intermediate4-(4-bromophenyl)-1,2,3,6-tetrahydropyridine (17) in an embodiment ofthe invention (FIG. 2).

4-(4-Bromophenyl)piperidin-4-ol 16 (1.0 g, 3.90 mmol) was carefullyadded to CF₃COOH (2.99 mL, 39 mmol), and the resulting mixture washeated at 90° C. for 3 h. After cooling, the solvent was removed undervacuum to give the title product as a white solid (0.90 g, 97%). ¹H NMR(400 MHz, Methanol-d₄) δ 7.54 (d, J=8.6 Hz, 2H), 7.41 (d, J=8.6 Hz, 2H),6.34-6.00 (m, 1H), 3.85 (dd, J=2.7 Hz, 2H), 3.48 (t, J=6.1 Hz, 2H),2.93-2.60 (m, 2H). 13C NMR (100 MHz, MeOD) δ: 23.3, 40.7, 42.0, 116.4,121.7, 126.6, 131.4, 134.6, 138.1. m/z (ESI, MH⁺) 239. ESI-HRMS (MH⁺)calcd. for C₁₁H₁₂BrN 238.0231, found 238.0230.

Example 5

This example demonstrates the synthesis of the intermediate4-(4-bromophenyl)piperidine (18) in an embodiment of the invention (FIG.2).

To a solution of 4-(4-bromophenyl)-1,2,3,6-tetrahydropyridine 17 (0.90g, 3.78 mmol) in dry MeOH (20 mL) and Et₃N (2 ml) was added Rh/Ccatalyst (0.060 g, J. Bishop & Co. Platinum). The resulting reactionmixture was stirred at room temperature in a hydrogen atmosphere (100psi) for 24 h. The mixture was filtered through a cake of CELITE™ (SigmaAldrich, St. Louis, Mo.), and the filtrate was concentrated to give thetitle compound as a white solid (0.91 g, 98%). TH NMR (400 MHz, MeOD) &:1.55-1.59 (2H, m). 1.61-1.70 (2H, m), 2.55-2.56 (1H, m), 2.64-2.70 (2H,m), 3.09-3.06 (2H, m), 713 (2H, J=8.0, d), 7.31 (2H, J=8.0, d). ¹³C NMR(100 MHz, MeOD) δ: 32.7, 41.4, 45.6, 119.4, 126.4, 128.2, 128.5, 131.2,145.3. m/z (EST, MH⁺) 241.

Example 6

This example demonstrates the synthesis of the intermediate methyl5-((tert-butoxycarbonyl)amino)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(19) in an embodiment of the invention (FIG. 2).

A suspension of 15 (0.514 g, 01.3 mmol), K₂CO₃ (0.565 g, 4.0 mmol) indry DME (10 mL) was stirred for 15 min. 18 (0.555 g, 1.9 mmol) was addedand the yellow suspension was degassed with N₂ for 40 min. Then,Pd(Ph₃P)₄ (0.078 g, 0.068 mmol) was added to the resulting mixture whileflushing N₂ for an additional 5 min. The reaction was heated at 85° C.for 7 h; after cooling the mixture was filtered through CELITE™ (SigmaAldrich, St. Louis, Mo.), and the solvent was removed under vacuum. Theresidue was purified by silica gel chromatography using as eluant DCM:MeOH: Et₃N (9:1:0.1) to afford to the title compound as a white solid(0.548 g, 70%). ¹H NMR (400 MHz, CDCl₃) δ:1.47 (9H, s), 1.85-1.90 (4H,m), 2.65-2.68 (1H, m), 2.78-2.83 (2H, m), 3.28-3.37 (2H, m), 3.85 (3H,s), 6.67 (1H, br s), 7.04-7.06 (1H, m), 7.24 (2H, J=8, d), 7.52 (2H,J=8, d), 7.84 (2H, m). ¹³C NMR (100 MHz, CDCl₃) δ: 28.4, 29.7, 33.8,42.3, 42.7, 46.7, 52.2, 61.1, 118.0, 122.6, 126.2, 126.8, 127.3, 128.4,128.6, 131.5 137.9, 139.3, 142.1, 146.0, 152.8, 166.9. m/z (ESI, MH⁺)411. ESI-HRMS (MH⁺) calcd. for C₂₄H₃₀N₂O₄ 411.2284, found 411.2285.

Example 7

This example demonstrates the synthesis of the intermediate methyl5-((tert-butoxycarbonyl)amino)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(20) in an embodiment of the invention (FIG. 2).

To a suspension of 19 (0.538 g, 1.3 mmol) in dry Et₂O (2.5 mL) at 0° C.and N₂ atmosphere, Et₃N (0.43 ml, 3.1 mmol) and TFAA (0.34 ml, 2.4 mmol)were added, and the resulting mixture was stirred for 1 h. The organicsolvent was removed under vacuum and the resulting orange oil was usedin the next step without any further purification. m/z (ESI, MH⁺) 507.ESI-HRMS (MH⁺) calcd. for C₂₆H₂₉F3N₂O₅ 529.1926, found 529.1935.

Example 8

This example demonstrates the synthesis of the intermediate methyl5-amino-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(21) in an embodiment of the invention (FIG. 2).

To a solution of 20 (0.538 g, 1.06 mmol) in DCM (12 mL), TFA (2.44 ml,31.8 mmol) was added and the resulting mixture was stirred overnight. Asaturated solution of NaHCO₃ was gradually added to get neutral pH andthe aqueous layer was extracted with DCM (3×30 mL). The collectedorganic fractions were dried over Na₂SO₄, filtered and the solvent wasremoved under vacuum. The residue was purified by silica gelchromatography using as eluant hexane/EtOAc (60:40) to give a yellow oil(0.371 g, 70%). (ESI, MH⁺) 407. ¹H NMR (400 MHz, Chloroform-d) δ 7.65(t, J=1.5 Hz, 1H), 7.55 (d, J=8.3 Hz, 2H), 7.33 (t, J=1.5 Hz, 1H), 7.26(d, J=8.2 Hz, 2H), 7.06 (t, J=1.5 Hz, 1H), 4.72 (dd, 1H), 4.16 (d,J=13.7 Hz, 1H), 3.91 (s, 3H), 3.27 (td, 1H), 3.01-2.72 (m, 2H), 2.02 (d,J=12.9 Hz, 2H), 1.75 (qd, J=4.0, 12.7 Hz, 2H). ¹³C NMR (100 MHz, CDCl₃)δ: 32.6, 33.6, 42.1, 44.2, 46.4, 52.1, 114.8, 117.8, 118.7, 120.3,121.2, 122.7, 127.5 (t, J_(C-F)=41 Hz), 131.6, 138.9, 139.1 142.1,143.7, 146.7, 167.1. (ESI, MH⁺) 407. ESI-HRMS (MH⁺) calcd. forC₂₁H₂₁F₃N₂O₃ 407.1583, found 407.1576.

Example 9

This example demonstrates the synthesis of the intermediate methyl5-azido-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(22) in an embodiment of the invention (FIG. 2).

To a solution of 21 (0.121 g, 0.29 mmol) in a 3:7 mixture of H₂O and ACN(10 ml), pTs-OH (0.509 g, 2.6 mmol) was added, and the mixture wasstirred for 5 min. NaNO₂ (0.184 g, 2.6 mmol) was then added, and theyellow solution was stirred at room temperature. The course of thereaction was followed using TLC (Hex/EtOAc 60:40), and the reaction wasallowed to continue until the starting material disappeared. NaN₃ (0.030g, 0.47 mmol) was added at room temperature, and the reaction mixturewas stirred overnight. Et₂O was added and the phases were separated. Theorganic phase was dried over Na₂SO₄, filtered and concentrated undervacuum to give an orange oil that was purified by silica gelchromatography using as eluant hexane/EtOAc (70:30) to afford the titlecompound as a yellow oil (0.107 g, 83%). ¹H NMR (400 MHz, chloroform-d)δ 8.03 (t, J=1.6 Hz, 1H), 7.68 (t, J=1.6 Hz, 1H), 7.56 (d, J=8.3 Hz,2H), 7.37 (t, J=2.0 Hz, 1H), 7.30 (d, J=8.2 Hz, 2H), 4.80-4.63 (m, 1H),4.17 (dd, J=4.0, 14.2 Hz, 1H), 3.96 (s, 3H), 3.27 (td, J=2.4, 13.3 Hz,1H), 2.99-2.76 (m, 2H), 2.02 (d, J=15.0 Hz, 2H), 1.76 (qd, J=4.2, 12.8Hz, 2H). ¹³C NMR (101 MHz, Chloroform-d) δ 166.6, H, 155.9, H, 144.9, H,143.2, H, 141.5, H, 138.1, H, 132.7, H, 127.8, H, 125.1, H, 122.1, H,118.7, H, 52.9, H, 46.8, H, 44.6, H, 42.5, H, 34.0, H, 33.0, H.

Example 10

This example demonstrates a general procedure for the clickcycloaddition reaction for the synthesis of compounds or intermediatesin an embodiment of the invention (FIG. 2).

To a solution of aryl azide (1 eq) and aryl alkyne (1.5 eq) in 2 mL oftetrahydrofuran (THF):water (1:1), sodium ascorbate (freshly prepared 1M aqueous solution) and CuSO₄ (0.5 eq) were sequentially added. Theresulting reaction was vigorously stirred for 12 h at room temperature.The reaction mixture was then concentrated in vacuo and purified byflash chromatography (hexane:ethyl acetate=6:4).

Methyl5-(4-phenyl-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23a). Yellow solid, m/z (ESI, MH⁺) 535; ESI-HRMS (MH⁺) calcd. forC₂₉H₂₆F₃N₄O₃ 535.1952, found 535.1957.

Methyl5-(4-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23b). Orange sticky solid, m/z (ESI, MH⁺) 603; ESI-HRMS (MH⁺) calcd.for C₃₀H₂₅F₆N₄O₃ 603.1825, found 603.1831.

Methyl 5-(4-(4-(ethylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23c). Yellow solid 2.8 mg (43%), m/z (ESI, MH⁺) 563.2; ESI-HRMS (MH⁺)calcd. for C₃₁H₃₀F₃N₄O₃ 563.2270, found 563.2274.

Methyl5-(4-(4-(hydroxymethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23d). Yellow solid 1.6 mg (25%), m/z (ESI, M+H⁺) 565.2; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₈F₃N₄O₄ 565.2063, found 565.2068.

Methyl5-(4-(3-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23e). Yellow solid 4 mg (80%), m/z (ESI, M+H⁺) 565.1; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₈F₃N₄O₄ 565.2063, found 565.2056.

Methyl5-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23f). Yellow solid 1.7 mg(27%), m/z (ESI, M+H⁺) 550.2; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₇F₃N₅O₃550.2066, found 550.2075.

Methyl5-(4-(4-chlorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23g). Yellow solid 3.6 mg(55%), m/z (ESI, M+H⁺) 569.2; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅³⁵ClF₃N₄O₃ 569.1567, found 569.1561.

Methyl5-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23h). Yellow solid 3.8 mg(54%), m/z (ESI, M+H⁺) 613.1; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅⁷⁹BrF₃N₄O₃ 613.1062, found 613.1057.

Methyl5-(4-(thiophen-3-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23i). Yellow solid 4.2 mg (67%), m/z (ESI, M+H⁺) 541.1.

Methyl 5-(4-(thiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23j). Yellow solid 1.6 mg (26%), m/z (ESI, M+H⁺) 541.2; ESI-HRMS (M+H⁺)calcd. for C₂₇H₂₄F₃N₄O₃ ³²S 541.1521, found 541.1523.

Methyl5-(4-(5-chlorothiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)-piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23k). Brown solid, m/z (ESI, MH⁺) 575; ESI-HRMS (MH⁺) calcd. forC₂₇H₂₃F₃N₄O₃SCl 575.1131, found 575.1132.

Methyl 5-(4-(5-bromothiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)-piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23l). Orange solid 5.1 mg (72%), m/z (ESI, MH⁺) 619.0; ESI-HRMS (MH⁺)calcd. for C₂₇H₂₃F₃N₄O₃S⁷⁹Br 619.0626, found 619.0618.

Methyl5-(4-(4-methylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23o). Yellow solid 4 mg (80%), m/z (ESI, M+H⁺) 549.2; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₈F₃N₄O₄ 549.2114, found 549.2119.

Methyl5-(4-(4-isopropylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(24q). Colorless oil 4 mg (60%), m/z (ESI, M+H⁺) 577.2; ESI-HRMS (M+H⁺)calcd. for C₃₂H₃₂F₃N₄O₃ 577.2427, found 577.2417.

Methyl5-(4-(4-(tert-butyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23r). Yellow solid 5.5 mg (81%), m/z (ESI, M+H⁺) 591.2; ESI-HRMS (M+H⁺)calcd. for C₃₃H₃₄F₃N₄O₃ 591.2583, found 591.2589.

Methyl 5-(4-(3-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23t). Yellow solid 6.15 mg (89%), m/z (ESI, M+H⁺) 603.2.

Methyl5-(4-(4-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23u). Yellow solid 3.2 mg(49%), m/z (ESI, M+H⁺) 565.2; ESI-HRMS (M+H⁺) calcd. for C₃₀H₂₈F₃N₄O₄565.2063, found 565.2062.

Methyl5-(4-(4-(pentyloxy)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23v). Pale yellow oil 5.7 mg (81%), m/z (ESI, M+H⁺) 621.2; ESI-HRMS(M+H⁺) calcd. for C₃₄H₃₆F₃N₄O₄ 621.2689, found 621.2687.

Methyl5-(4-(2-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23w). Yellow solid 6.5 mg (99%), m/z (ESI, M+H⁺) 565.1; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₈F₃N₄O₄ 565.2063, found 565.2061.

Methyl 5-(4-(4-(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23x). Yellow solid 3.4 mg (48%), m/z (ESI, M+H⁺) 619.1; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₅F₆N₄O₄ 619.1780, found 619.1784.

Methyl 5-(4-(3-(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23y). Yellow solid 6 mg (84%), m/z (ESI, M+H⁺) 619.2; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₅F₆N₄O₄ 619.1780, found 619.1778.

Methyl5-(4-(3-chlorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23z). Yellow solid 3.5 mg(54%), m/z (ESI, M+H⁺) 569.2; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅³⁵ClF₃N₄O₃ 569.1567, found 569.1570.

Methyl5-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23aa). Yellow solid 3.9mg (61%), m/z (ESI, M+H⁺) 553.1; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅F₄N₄O₃553.1863, found 553.1855.

Methyl5-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23bb). Yellow solid 4.5mg (77%), m/z (ESI, M+H⁺) 553.2; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅F₄N₄O₃553.1863, found 553.1872.

Methyl5-(4-(2-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23cc). Yellow solid 4.9mg (71%), m/z (ESI, M+H⁺) 553.2; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₅F₄N₄O₃553.1863, found 553.1866.

Methyl5-(4-(3,4-difluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23dd). Pale yellow solid 4.23 mg (65%), m/z (ESI, M+H⁺) 571.1; ESI-HRMS(M+H⁺) calcd. for C₂₉H₂₄F₅N₄O₃ 571.1769, found 571.1758.

Methyl5-(4-(3,5-difluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23ee). Yellow solid 4.2 mg (63%), m/z (ESI, M+H⁺) 571.1; ESI-HRMS(M+H⁺) calcd. for C₂₉H₂₄F₄N₅O₃ 571.1769, found 571.1762.

Methyl5-(4-(4-cyanophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23ff). Yellow solid 6 mg (94%), m/z (ESI, M+H⁺) 560.2; ESI-HRMS (M+H⁺)calcd. for C₃₀H₂₅F₄N₅O₃ 560.1909, found 560.1913.

Methyl5-(4-(4-acetylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23hh). Yellow solid 4.45 mg (67%), m/z (ESI, M+H⁺) 577.2; ESI-HRMS(M+H⁺) calcd. for C₃₁H₂₈F₃N₄O₄ 577.2063, found 577.2056.

Methyl5-(4-(3,4-dimethoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23ii). Yellow solid 5.65 mg (83%), m/z (ESI, M+H⁺) 595.2; ESI-HRMS(M+H⁺) calcd. for C₃₁H₃₀F₃N₄O₅ 595.2168, found 595.2173.

Methyl5-(4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23jj). Yellow solid 5.15mg (81%), m/z (ESI, M+H⁺) 551.1; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₆F₃N₄O₄551.1906, found 551.1901.

Methyl5-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate (23kk). Yellow solid 2.4mg (38%), m/z (ESI, M+H⁺) 550.1; ESI-HRMS (M+H⁺) calcd. for C₂₉H₂₇F₃N₅O₃550.2066, found 550.2056.

Methyl 5-(4-(pyridin-4-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23ll). Yellow solid 2.3 mg (37%), m/z (ESI, M+H⁺) 536.1; ESI-HRMS(M+H⁺) calcd. for C₂₈H₂₅F₃N₅O₃ 536.1909, found 536.1913.

Methyl5-(4-(pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23 mm). Yellow solid 3.5 mg (57%).

Methyl5-(4-(furan-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23nn). Yellow solid 3 mg (50%), m/z (ESI, M+H⁺) 525.1; ESI-HRMS (M+H⁺)calcd. for C₂₇H₂₄F₃N₄O₄ 525.1750, found 525.1744.

Methyl 5-(4-(benzofuran-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23oo). Brown solid 4 mg (60%), m/z (ESI, M+H⁺) 575.2; ESI-HRMS (M+H⁺)calcd. for C₃₁H₂₆F₃N₄O₄ 575.1906, found 575.1909.

Methyl5-(4-(thiazol-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23pp). Yellow solid 4.6 mg (74%), m/z (ESI, M+H⁺) 542.1; ESI-HRMS(M+H⁺) calcd. for C₂₆H₂₃F₃N₅O₃ ³²S 542.1474, found 542.1465.

Methyl5-(4-cyclohexyl-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23qq). Yellow solid 5.4 mg (87%), m/z (ESI, M+H⁺) 541.2; ESI-HRMS(M+H⁺) calcd. for C₂₉H₃₂F₃N₄O₃ 541.2427, found 541.2419.

Methyl5-(4-cyclopropyl-1H-1,2,3-triazol-1-yl)-4′-(1-(2,2,2-trifluoroacetyl)piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylate(23rr). Yellow solid 5.7 mg (99%), m/z (ESI, M+H⁺) 499.2; ESI-HRMS(M+H⁺) calcd. for C₂₆H₂₆F₃N₄O₃ 499.1957, found 499.1959.

Example 11

This example demonstrates the synthesis of5-(4-phenyl-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24a) in an embodiment of the invention (FIG. 2).

To a solution of 23a (9 mg, 17 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(9.5 mg, 170 μmol) was added and the resulting mixture was heated at 50°C. for 12 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=25.7 min). The product wasobtained as a white solid by lyophilization (1.9 mg, 26%). (ESI, MH⁺)425; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₅N₄O₂ 425.1978, found 425.1978, HPLCpurity 97.1% (Rt=8.9 min). ¹H NMR (400 MHz, MeOD) δ:1.77-1.80 (3H, m),1.94-1.97 (2H, m), 2.90-2.96 (4H, m), 7.31-7.35 (3H, m), 7.39-7.43 (2H,m), 7.68 (2H, J=8 Hz, d), 7.88 (2H, J=8 Hz, d), 8.29 (2H, J=8 Hz, d),8.94 (1H, s).

Example 12

This example demonstrates the synthesis of5-(4-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24b) in an embodiment of the invention (FIG. 2).

To a solution of 23b (6.3 mg, 10 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=30.2 min). The productwas obtained as a white solid by lyophilization (2.1 mg, 42%). (ESI,MH⁺) 493; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₄F₃N₄O₂ 493.1851, found493.1851; HPLC purity 96.9% (R_(t)=10.1 min). ¹H NMR (400 MHz, MeOD)δ:1.79-1.84 (2H, m), 1.98-2.04 (2H, m), 3.01-3.06 (4H, m), 7.45 (2H, J=8Hz, d), 7.69-7.73 (4H, m), 8.10 (2H, J=8 Hz, d), 8.43 (2H, J=8 Hz, d),8.99 (1H, s).

Example 13

This example demonstrates the synthesis of5-(4-(4-ethylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24c) in an embodiment of the invention (FIG. 2).

To a solution of 23c (2.8 mg, 5 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=27.0 min). The productwas obtained as a white solid by lyophilization (2.4 mg, 67%). (ESI,MH⁺) 453.2; ESI-HRMS (MH⁺) calcd. for C₂₈H₂₉N₄O₂ 453.2291, found453.2294; HPLC purity 98.6% (R_(t)=11.5 min).

Example 14

This example demonstrates the synthesis of5-(4-(4-(hydroxymethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24d) in an embodiment of the invention (FIG. 2).

To a solution of 23d (1.6 mg, 2.8 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=23.3 min). The productwas obtained as a white solid by lyophilization (1.4 mg, 99%). (ESI,MH⁺) 455.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₇N₄O₃ 455.2083, found455.2081; HPLC purity 96.3% (R_(t)=8.42 min).

Example 15

This example demonstrates the synthesis of5-(4-(3-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24e) in an embodiment of the invention (FIG. 2).

To a solution of 23e (4 mg, 7 μmol) in 2 mL of MeOH: H₂O (1:1), KOH (5.6mg, 100 μmol) was added, and the resulting mixture was heated at 50° C.for 12 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=23.2 min). The product wasobtained as a white solid by lyophilization (2.4 mg, 67%). (ESI, MH⁺)455.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₇N₄O₃ 455.2083, found 455.2083;HPLC purity 99.8% (R_(t)=10.1 min). ¹H NMR (500 MHz, DMSO-d₆): δ(ppm)=9.52 (s, 1H, CH_(triazole)), 8.42 (s, 1H), 8.33 (s, 1H), 8.19 (s,1H), 7.78 (d, J=7.8 Hz, 2H), 7.63-7.52 (m, 1H), 7.42 (m, 3H), 6.95 (d,J=8.1 Hz, 1H), 6.81 (s, 1H), 3.84 (s, 3H, CH₃), 2.90 (m, 3H), 1.91 (m,5H). Proton signals for 2-CH_(piperidine), 6-CH_(piperidine) and3-CH_(piperidine), 5-CH_(piperidine) are hidden under the H₂O andDMSO-d₆ signals. ¹³C NMR (500 MHz, DMSO-d₆): δ (ppm)=172.8 (1C), 168.7(1C), 160.3 (1C), 147.7 (1C), 145.9 (1C), 141.3 (1C), 137.8 (1C), 137.3(1C), 132.2 (1C), 130.7 (2C), 127.9 (2C), 127.5 (2C), 127.4 (1C), 120.5(1C), 119.5 (1C), 119.1 (2C), 118.1 (1C), 114.5 (1C), 111.0 (2C), 55.7(2C), 44.6 (2C), 30.8 (1C), 29.5 (1C).

Example 16

This example demonstrates the synthesis of5-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24f) in an embodiment of the invention (FIG. 2).

To a solution of 23f (3.3 mg, 6 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=17.4 min). The productwas obtained as a white solid by lyophilization (2.84 mg, 95%). (ESI,MH⁺) 440.2; HPLC purity 96% (R_(t)=8.38 min).

Example 17

This example demonstrates the synthesis of5-(4-(4-chlorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24g) in an embodiment of the invention (FIG. 2).

To a solution of 23g (3.6 mg, 6.3 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=22.7 min). The productwas obtained as a white solid by lyophilization (3.3 mg, 99%). (ESI,MH⁺) 459.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄ ³⁵ClN₄O₂ 459.1588, found459.1588; HPLC purity 98.4% (R_(t)=11.4 min). ¹H NMR (500 MHz, DMSO-d₆):δ (ppm)=9.49 (s, 1H), 8.34 (s, 1H), 8.24 (s, 1H), 8.10 (s, 1H), 8.01 (d,J=8.6 Hz, 2H), 7.72 (d, J=8.4 Hz, 2H), 7.58 (d, J=8.6 Hz, 2H), 7.38 (d,J=8.4 Hz, 2H), 3.08 (d, J=12.2 Hz, 2H), 2.69-2.59 (m, 3H), 1.81-1.70 (m,5H), 1.60 (qd, J=3.8, 12.4 Hz, 2H). ¹³C NMR (500 MHz, DMSO-d₆): δ(ppm)=173.6 (1C), 167.6 (1C), 145.0 (1C), 146.6 (1C), 144.2 (1C), 141.1(2C), 137.5 (1C), 136.9 (2C), 133.0 (2C), 129.7 (2C), 129.5 (2C), 127.8(2C), 127.5 (2C), 127.3 (2C), 120.6 (2C), 119.6 (2C), 118.3 (2C), 49.0(1C), 46.5 (2C), 42.2 (1C), 33.9 (2C), 23.6 (1C).

Example 18

This example demonstrates the synthesis of5-(4-(4-bromophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24h) in an embodiment of the invention (FIG. 2).

A solution of 23h (3.8 mg, 6.2 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=24.2 min). Theproduct was obtained as a white acetate salt by lyophilization (3.5 mg,99%). (ESI, MH⁺) 503.1; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄N₄O₂ ⁷⁹Br503.1083, found 503.1080.

Example 19

This example demonstrates the synthesis of5-(4-(thiophen-3-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24i) in anembodiment of the invention (FIG. 2).

A solution of 23i (4.2 mg, 7.7 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=22.3 min). Theproduct was obtained as a white acetate salt by lyophilization (3.77 mg,99%). (ESI, MH⁺) 431.1; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₃N₄O₂ ³²S431.1542, found 431.1548, HPLC purity 99% (Rt=9.7 min).

Example 20

This example demonstrates the synthesis of5-(4-(thiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24j) in an embodiment of the invention (FIG. 2).

A solution of 23j (2.6 mg, 5 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=21.8 min). Theproduct was obtained as a white acetate salt by lyophilization (2.2 mg,99%). (ESI, MH⁺) 431.2; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₃N₄O₂ ³²S431.1542, found 431.1538, HPLC purity 97.1% (Rt=8.9 min).

Example 21

This example demonstrates the synthesis of5-(4-(5-chlorothiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24k) in an embodiment of the invention (FIG. 2).

To a solution of 23k (8.3 mg, 14 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=28.6 min). The product wasobtained as a white solid by lyophilization (2.8 mg, 42%). (ESI, MH⁺)465; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₂N₄O₂SCl 465.1152, found 465.1151;HPLC purity 97.7% (Rt=10.6 min).

Example 22

This example demonstrates the synthesis of5-(4-(5-bromothiophen-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24l) in an embodiment of the invention (FIG. 2).

To a solution of 23l (5.1 mg, 8.2 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=27.2 min). The product wasobtained as a white solid by lyophilization (2.31 mg, 50%). (ESI, MH⁺)509.1; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₂N₄O₂SBr 509.0647, found 509.0648;HPLC purity 97.7% (Rt=11.8 min).

Example 23

This example demonstrates the synthesis of5-(4-(4-methylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24o) in anembodiment of the invention (FIG. 2).

To a solution of 23o (6 mg, 10 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=24.4 min). The product wasobtained as a white solid by lyophilization (1.9 mg, 43%).

Example 24

This example demonstrates the synthesis of5-(4-(4-propylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24p) in anembodiment of the invention (FIG. 2).

To a solution of 23p (5 mg, 8.7 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=23.7 min). The product wasobtained as a white solid by lyophilization (4.5 mg, 98%). (ESI, MH⁺)467.2; ESI-HRMS (MH⁺) calcd. for C₂₉H₃₁N₄O₂ 467.2447, found 467.2454.

Example 25

This example demonstrates the synthesis of5-(4-(4-isopropylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24q) in anembodiment of the invention (FIG. 2).

To a solution of 23q (4 mg, 6.9 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=18.1 min). The product wasobtained as a white solid by lyophilization (3.55 mg, 97%). (ESI, MH⁺)467.2; ESI-HRMS (MH⁺) calcd. for C₂₉H₃₁N₄O₂ 467.2447, found 467.2440.

Example 26

This example demonstrates the synthesis of5-(4-(4-(tert-butyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24r) in an embodiment of the invention (FIG. 2).

To a solution of 23r (5.5 mg, 9.3 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=31.1 min). The product wasobtained as a white solid by lyophilization (5.03 mg, 99%). (ESI, MH⁺)481.3; ESI-HRMS (MH⁺) calcd. for C₃₀H₃₃N₄O₂ 481.2604, found 481.2597.HPLC purity 98.2% (R_(t)=12.9 min).

Example 27

This example demonstrates the synthesis of5-(4-(3-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24t) in anembodiment of the invention (FIG. 2).

To a solution of 23t (6.15 mg, 10 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=27.6 min). The product wasobtained as a white solid by lyophilization (2.53 mg, 46%). (ESI, MH⁺)493.1; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₄N₄O₂F₃ 493.1851, found 493.1851.HPLC purity 99% (R_(t)=11.8 min).

Example 28

This example demonstrates the synthesis of5-(4-(4-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24u) in an embodiment of the invention (FIG. 2).

To a solution of 23u (3.2 mg, 5.7 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=23.0 min). The product wasobtained as a white solid by lyophilization (2.14 mg, 73%). (ESI, MH⁺)455.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₇N₄O₃ 455.2083, found 455.2086.HPLC purity 97% (R_(t)=10.14 min).

Example 29

This example demonstrates the synthesis of5-(4-(4-(pentyloxy)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24v) in an embodiment of the invention (FIG. 2).

To a solution of 23v (5.8 mg, 9.3 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=34.1 min). The product wasobtained as a white solid by lyophilization (5.3 mg, 99%). (ESI, MH⁺)511.3; ESI-HRMS (MH⁺) calcd. for C₃₁H₃₅N₄O₃ 511.2709, found 511.2703.

Example 30

This example demonstrates the synthesis of5-(4-(2-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24w) in an embodiment of the invention (FIG. 2).

To a solution of 23w (6.5 mg, 11.5 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(7.9 mg, 140 μmol) was added and the resulting mixture was heated at 60°C. for 6 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=23.0 min). The product wasobtained as a white solid by lyophilization (3.05 mg, 52%). (ESI, MH⁺)455.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₇N₄O₃ 455.2083, found 455.2086.HPLC purity 98% (R_(t)=10.44 min).

Example 31

This example demonstrates the synthesis of5-(4-(3-(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24y) in an embodiment of the invention (FIG. 2).

A solution of 23y (6 mg, 9.7 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=29.2 min). Theproduct was obtained as a white acetate salt by lyophilization (3.63 mg,66%). (ESI, MH⁺) 509.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₄N₄O₃F₃ 509.1801,found 509.1798.

Example 32

This example demonstrates the synthesis of5-(4-(3-chlorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24z) in an embodiment of the invention (FIG. 2).

To a solution of 23z (3.5 mg, 6.2 μmol) in 2 mL of MeOH: H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=21.2 min). The productwas obtained as a white solid by lyophilization (2.2 mg, 68%). (ESI,MH⁺) 459.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄ ³⁵ClN₄O₂ 459.1588, found459.1583; HPLC purity 98% (R_(t)=11.1 min).

Example 33

This example demonstrates the synthesis of5-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24aa) in an embodiment of the invention (FIG. 2).

A solution of 23aa (3.87 mg, 7 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=20.6 min). Theproduct was obtained as a salt by lyophilization (3.06 mg, 99%). (ESI,MH⁺) 443.1; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄FN₄O₂ 443.1883, found443.1890, HPLC purity 96% (R_(t)=10.8 min).

Example 34

This example demonstrates the synthesis of5-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24bb) in an embodiment of the invention (FIG. 2).

A solution of 23bb (4.5 mg, 8.2 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=20.5 min).The product was obtained as a white acetate salt by lyophilization (3.4mg, 84%). (ESI, MH⁺) 443.1; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄FN₄O₂443.1883, found 443.1884, HPLC purity 98.2% (Rt=10.5 min). ¹H NMR (500MHz, DMSO-d6): δ (ppm)=9.57 (s, 1H), 8.37 (s, 1H), 8.24 (s, 1H), 8.11(s, 1H), 7.85 (t, J=8.1 Hz, 1H), 7.80 (t, J=8.1 Hz, 1H), 7.72 (d, J=8.3Hz, 2H), 7.56 (p, J=8.1 Hz, 1H), 7.38 (d, J=8.3 Hz, 2H), 3.10 (t, J=8.8Hz, 2H), 2.75-2.59 (m, 3H), 1.81-1.72 (m, 5H), 1.63 (m, 2H).

Example 35

This example demonstrates the synthesis of5-(4-(2-fluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24cc) in an embodiment of the invention (FIG. 2).

A solution of 23cc (4.9 mg, 8.8 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=24.3 min).The product was obtained as a white acetate salt by lyophilization (2.64mg, 60%). (ESI, MH⁺) 443.1; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₄FN₄O₂443.1887, found 443.1884.

Example 36

This example demonstrates the synthesis of5-(4-(3,4-difluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24dd) in an embodiment of the invention (FIG. 2).

A solution of 23dd (4.2 mg, 7.4 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=21.1 min).The product was obtained as a white acetate salt by lyophilization (3.85mg, 99%). (ESI, MH⁺) 461.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₃F₂N₄O₂461.1789, found 461.1790.

Example 37

This example demonstrates the synthesis of5-(4-(3,5-difluorophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24ee) in an embodiment of the invention (FIG. 2).

A solution of 23ee (4.16 mg, 7.3 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=22.2 min).The product was obtained as a white acetate salt by lyophilization (1.24mg, 33%). (ESI, MH⁺) 461.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₃F₂N₄O₂461.1789, found 461.1782.

Example 38

This example demonstrates the synthesis of5-(4-(4-carboxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24ff) in an embodiment of the invention (FIG. 2).

A solution of 23ff (6 mg, 10 μmol) in of 2 mL MeOH:1.0 M KOH (1:1) washeated at 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=9.4 min). Theproduct was obtained as a white acetate salt by lyophilization, (ESI,MH⁺) 469.2; ESI-HRMS (MH⁺) calcd. for C₂₇H₂₅F₂N₄O₄ 469.1876, found469.1870.

Example 39

This example demonstrates the synthesis of5-(4-(3-carboxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24gg) in an embodiment of the invention (FIG. 2).

To a solution of aryl azide (5 mg, 11.5 μmol, 1 eq) and 3-ethynylbenzoicacid (2.5 mg, 17 μmol, 1.5 eq) in 2 mL of THF:water (1:1), sodiumascorbate (17 μmol, freshly prepared 1 M aqueous solution, 1.5 eq) andCuSO₄ (1.4 mg, 6 μmol, 0.5 eq) were sequentially added. The resultingreaction was vigorously stirred for 12 h at room temperature. Thereaction mixture was then concentrated in vacuo and added to a solutionof 2 mL MeOH:1.0 M KOH (1:1), the mixture was heated at 50° C. for 12 h.After removing the solvents under vacuum, the mixture was purified bypreparative HPLC (Method B, R_(t)=10.4 min). The product was obtained asa white acetate salt by lyophilization (5.7 mg, 93%)., (ESI, MH⁺) 469.2;ESI-HRMS (MH⁺) calcd. for C₂₇H₂₅F₂N₄O₄ 469.1876, found 469.1884.

Example 40

This example demonstrates the synthesis of5-(4-(4-acetylphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24hh) in an embodiment of the invention (FIG. 2).

A solution of 23hh (4.5 mg, 7.7 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=19.4 min).The product was obtained as a white acetate salt by lyophilization (3.88mg, 95%). (ESI, MH⁺) 467.2; ESI-HRMS (MH⁺) calcd. for C₂₈H₂₇N₄O₃467.2083, found 467.2084, HPLC purity 96.6% (R_(t)=10.4 min).

Example 41

This example demonstrates the synthesis of5-(4-(3,4-dimethoxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24ii) in an embodiment of the invention (FIG. 2).

A solution of 23ii (5.65 mg, 9.5 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=13.5 min).The product was obtained as a white acetate salt by lyophilization (5.2mg, 99%). (ESI, MH⁺) 485.2; ESI-HRMS (MH⁺) calcd. for C₂₈H₂₉N₄O₄485.2189, found 485.2195.

Example 42

This example demonstrates the synthesis of5-(4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24jj) in an embodiment of the invention (FIG. 2).

A solution of 23jj (5.15 mg, 9.3 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=17.9 min).The product was obtained as a white acetate salt by lyophilization (4.7mg, 99%). (ESI, MH⁺) 441.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₅N₄O₃441.1927, found 441.1931, HPLC purity 96.2% (R_(t)=8.7 min).

Example 43

This example demonstrates the synthesis of5-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24kk) in an embodiment of the invention (FIG. 2).

To a solution of 23kk (2.4 mg, 4.3 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=17.4 min). The productwas obtained as a white solid by lyophilization (2.2 mg, 99%). (ESI,MH⁺) 440.2; ESI-HRMS (MH⁺) calcd. for C₂₆H₂₆N₅O₂ 440.2087, found440.2093.

Example 44

This example demonstrates the synthesis of5-(4-(pyridin-4-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24ll) in anembodiment of the invention (FIG. 2).

To a solution of 23ll (2.3 mg, 4.3 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=15.7 min). The productwas obtained as a white solid by lyophilization (2 mg, 94%). (ESI, MH⁺)426.1; ESI-HRMS (MH⁺) calcd. for C₂₅H₂₄N₅O₂ 426.1930, found 426.1933.

Example 45

This example demonstrates the synthesis of5-(4-(pyrazin-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24 mm) in anembodiment of the invention (FIG. 2).

To a solution of 23 mm (3.55 mg, 6.6 μmol) in 2 mL of MeOH:H₂O (1:1),KOH (5.6 mg, 100 μmol) was added, and the resulting mixture was heatedat 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=16.8 min). Theproduct was obtained as a white solid by lyophilization (2.81 mg, 88%).(ESI, MH⁺) 427.2; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₃N₆O₂ 427.1882, found427.1881.

Example 46

This example demonstrates the synthesis of5-(4-(furan-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24nn) in an embodiment of the invention (FIG. 2).

To a solution of 23nn (3 mg, 5.7 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=19.8 min). The productwas obtained as a white solid by lyophilization (2.28 mg, 84%). (ESI,MH⁺) 415.2; ESI-HRMS (MH⁺) calcd. for C₂₄H₂₃N₄O₃ 415.1770, found415.1767, HPLC purity 98% (R_(t)=9.2 min).

Example 47

This example demonstrates the synthesis of5-(4-(benzofuran-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24oo) in an embodiment of the invention (FIG. 2).

To a solution of 23oo (4 mg, 7 μmol) in 2 mL of MeOH:H₂O (1:1), KOH (5.6mg, 100 μmol) was added, and the resulting mixture was heated at 50° C.for 12 h. After removing the solvents under vacuum, the mixture waspurified by preparative HPLC (Method B, R_(t)=21.9 min). The product wasobtained as a white solid by lyophilization, (ESI, MH⁺) 465.2; ESI-HRMS(MH⁺) calcd. for C₂₈H₂₅N₄O₃ 465.1927, found 465.1936.

Example 48

This example demonstrates the synthesis of5-(4-(thiazol-2-yl)-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylic acid (24pp) in anembodiment of the invention (FIG. 2).

A solution of 23pp (4.6 mg, 8.5 μmol) in of 2 mL MeOH:1.0 M KOH (1:1)was heated at 50° C. for 12 h. After removing the solvents under vacuum,the mixture was purified by preparative HPLC (Method B, R_(t)=18.7 min).The product was obtained as a white acetate salt by lyophilization (4.18mg, 99%). (ESI, MH⁺) 431.2; ESI-HRMS (MH⁺) calcd. for C₂₃H₂₂N₅O₂ ³²S432.1494, found 432.1494, HPLC purity 96.4% (R_(t)=9.1 min).

Example 49

This example demonstrates the synthesis of5-(4-cyclohexyl-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24qq) in an embodiment of the invention (FIG. 2).

To a solution of 23qq (5.4 mg, 9.9 μmol) in 2 mL of MeOH:H₂O (1:1), KOH(5.6 mg, 100 μmol) was added, and the resulting mixture was heated at50° C. for 12 h. After removing the solvents under vacuum, the mixturewas purified by preparative HPLC (Method B, R_(t)=21.2 min). The productwas obtained as a white solid by lyophilization (2.65 mg, 55%). (ESI,MH⁺) 431.3; ESI-HRMS (MH⁺) calcd. for C₂₆H₃₁N₄O₂ 431.2447, found431.2444. HPLC purity 97% (R_(t)=10.4 min).

Example 50

This example demonstrates the synthesis of5-(4-cyclopropyl-1H-1,2,3-triazol-1-yl)-4′-(piperidin-4-yl)-[1,1′-biphenyl]-3-carboxylicacid (24rr) in an embodiment of the invention (FIG. 2).

To a solution of 23rr (5.7 mg, 11.5 μmol) in 2 mL of MeOH: H₂O (1:1),KOH (5.6 mg, 100 μmol) was added, and the resulting mixture was heatedat 50° C. for 12 h. After removing the solvents under vacuum, themixture was purified by preparative HPLC (Method B, R_(t)=12.6 min). Theproduct was obtained as a white solid by lyophilization (5.1 mg, 99%).(ESI, MH⁺) 389.2; ESI-HRMS (MH⁺) calcd. for C₂₃H₂₅N₄O₂ 389.1978, found389.1971.

Example 51

This example demonstrates the synthesis of4′-(piperidin-4-yl)-5-((4-(trifluoromethyl)phenyl)ethynyl)-[1,1′-biphenyl]-3-carboxylicacid hydrochloride (44) in an embodiment of the invention (FIG. 3).

3-Hydroxy-5-iodobenzoic acid (38a, 264 mg, 1 mmol) was suspended inmethanol (3 mL), and the solution was cooled to 0° C. in an ice bath.Thionyl chloride (0.5 mL, 7 mmol) was added to the mixture over thecourse of 30 min at 0° C. The mixture was allowed to warm to roomtemperature and stirred for 16 h. The solvent was removed from theresulting light yellow solution under vacuum and the residue wasredissolved in dichloromethane (3 mL). The solution was washed withsaturated aqueous sodium bicarbonate solution (1 mL) and water (1 mL).The organic layer was dried with sodium sulfate and filtered through apad of silica gel eluting with additional volume of dichloromethane. Thecombined eluants were evaporated to dryness to provide 38b (92 mg, 33%).¹H NMR (400 MHz, CDCl₃) δ 7.94 (s, 1H), 7.54 (s, 1H), 7.44 (s, 1H), 5.68(s, 1H), 3.93 (s, 3H). ¹³C NMR (101 MHz, CDCl₃) δ 165.9, 156.3, 132.6,130.9, 129.2, 116.1, 93.9, 52.7. Positive ESI-MS (m/z) 279.

1-Ethynyl-4-(trifluoromethyl)benzene (39b, 102 mg, 0.6 mmol) was addedto a degassed suspension of 38b (110 mg, 0.4 mmol),bis(triphenylphosphine)palladium(II) dichloride (14 mg, 5 mol %) andcopper(I) iodide (4 mg, 5 mol %) and triethylamine (0.3 mL, mmol) inanhydrous DMF (6 mL) at 0° C. The reaction mixture was allowed to warmup to stirred until the complete consumption of 38b. The reactionmixture was quenched with water (25 mL) and the organic products wereextracted with ethyl acetate (3×5 mL). The combined extracts were washedwith water (2×5 mL), brine (5 mL), dried with sodium sulfate andevaporated to dryness. The residue of evaporation was subjected tocolumn chromatography (silica gel), eluting with chloroform/methanol90/10 (v/v) mixture, to provide 40 (103 mg, 81%). ¹H NMR (400 MHz,CDCl₃) δ 7.80 (t, J=1.38 Hz, 1H), 7.63 (s, 4H), 7.60 (dd, J=1.51, 2.51Hz, 1H), 7.24 (dd, J=1.51, 2.51 Hz, 1H), 6.06 (br. s., 1H), 3.95 (s,3H). ¹³C NMR (101 MHz, CDCl₃) δ 166.5, 155.9, 131.9, 131.7, 126.6,125.4, 125.3, 125.3, 124.2, 122.8, 117.1, 116.1, 90.3, 88.7, 52.6.Positive ESI-MS (m/z) 321.

Trifluoromethanesulfonic anhydride (54 μL, 0.32 mmol) was added to asolution of 40 (93 mg, 0.29 mmol) and triethylamine (61 μL, 0.43 mmol)in anhydrous dichloromethane (2 mL) at −20° C. under inert atmosphere.The reaction mixture was removed from cooling bath and left to stir at23° C. for 2.5 h. The solution was diluted with dichloromethane (3 mL),washed with water (1 mL), saturated aqueous sodium bicarbonate solution(1 mL), dried with sodium sulfate and evaporated to dryness undervacuum. The residue following evaporation was subjected to columnchromatography (silica gel), eluting with ethyl acetate/hexane 20/80(v/v) mixture. The combined fractions containing 41 were evaporated todryness to afford product (128 mg, 98%). ¹H NMR (400 MHz, CDCl₃) δ 8.25(t, J=1.38 Hz, 1H), 7.92 (dd, J=1.38, 2.38 Hz, 1H), 7.66 (s, 4H), 7.63(s, 1H), 3.99 (s, 3H). Positive ESI-MS (m/z) 453.

A mixture of triflate 41 (25 mg, 56 μmol), tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine-1-carboxylate(42, 22 mg, 56 μmol), tetrakis(triphenylphosphine)palladium(0) (127 mg,0.11 mmol), potassium carbonate (15 mg, 0.11 mmol) and DMF (1 mL) wasdegassed and heated to 90° C. under the atmosphere of inert gas for 8 h.After cooling to 23° C., the solvent was removed under vacuum. Theresidue was resuspended in ethyl acetate (3 mL), washed with water (2×1mL) and dried with sodium sulfate. Ethyl acetate was removed undervacuum, and the residue was subjected to column chromatography (silicagel), eluting with chloroform/methanol 95/5 (v/v) mixture to obtain thetitle compound 43 (21 mg, 67%). ¹H NMR (400 MHz, CDCl₃) δ 7.86 (s, 1H),7.56 (m, 4H), 7.46-7.49 (m, 1H), 7.29 (m, 5H), 5.36 (m, 2H), 3.94 (s,3H), 2.84 (br. s., 2H), 2.71 (t, J=12.05 Hz, 1H), 1.86 (m, 2H), 1.68 (m,2H), 1.50 (s, 9H). Positive ESI-MS (m/z) 564.

Lithium hydroxide (aqueous 0.5M, 70 μL, 25 μmol) was added to a solutionof 43 (13 mg, 23 μmol) in methanol (0.2 mL), and the mixture was heatedat reflux for 1.5 h. During this time 43 was completely consumed. Themixture was allowed to cool to 23° C. and acidified with hydrochloricacid (1M) until pH 1. The acidified mixture was stirred for additional 2h before solvents were removed under vacuum. The residue was subjectedto column chromatography (silica gel), eluting withchloroform/methanol/acetic acid 100/10/1 (v/v) mixture. Hydrochloricacid (1 M) was added to fractions containing the desired product and thesolvent was removed under vacuum to provide the desired product 44 as ahydrochloride salt (3.1 mg, 28%). ¹H NMR (400 MHz, DMSO-d₆) δ 8.24 (s,1H), 8.11 (t, J=1.63 Hz, 1H), 8.08 (s, 1H), 7.85 (s, 1H), 7.79 (td,J=1.51, 7.78 Hz, 1H), 7.73 (td, J=1.38, 7.53 Hz, 1H), 7.69 (d, J=8.03Hz, 2H), 7.41 (t, J=7.65 Hz, 1H), 7.35 (d, J=8.28 Hz, 2H), 3.26 (br. s.,2H), 3.10-3.19 (m, 2H), 2.87 (d, J=12.55 Hz, 3H), 2.42 (dt, J=2.64, 7.22Hz, 2H), 2.33 (td, J=1.79, 3.70 Hz, 1H). Positive ESI-MS (m/z) 450.

Example 52

The synthesis of fluorescent antagonist 4 as previously reported(Kiselev et al., ACS Chem. Biol. 2014, 9, 2833-2842) suffered from a lowyield in the final click cycloaddition step to link theazide-functionalized fluorophore and the alkyne-functionalizedpharmacophore. For the purpose of screening antagonist analogues, largerquantities of a fluorescent probe were needed. Therefore, an alternateroute to 4 was designed and also explored another fluorescent antagonistanalogue for possible use in screening. Given the unusually highaffinity of 4 and its low nonspecific character, an alternate synthesisof the same probe was developed, in addition to modifying its structure,in order to provide a sufficient supply of 4 for use in routine assays.See FIGS. 4A and 4B.

This example demonstrates the synthesis of the intermediatehex-5-yn-1-yl 4-methylbenzenesulfonate (26) in an embodiment of theinvention (FIG. 4A).

To a solution of hex-5-yn-1-ol 25 (0.84 mL, 7.64 mmol), triethylamine(1.28 mL, 9.17 mmol), and 4-(dimethylamino)pyridine (20 mg, 0.15 mmol)in dichloromethane (DCM) (25 mL) at 0° C. was added p-toluenesulfonylchloride (1.53 g, 8.02 mmol) in three portions. The reaction mixture wasbrought to room temperature and stirred for 15 h. Aqueous NaOH (1 N, 15mL) was added, and the mixture was vigorously stirred for 15 min at roomtemperature. The usual workup (DCM, brine) gave the title compound (1.68g, 88%) as a yellowish oil. ¹H NMR (400 MHz, CDCl₃) δ: 1.42-1.48 (2H,m), 1.65-1.68 (2H, m), 1.84 (1H, J=4, t), 2.05-2.09 (2H, m), 2.31 (3H,s), 3.96 (2H, J=6, t), 7.26 (2H, J=8.0, d), 7.69 (2H, J=8.0, d). ¹³C NMR(100 MHz, CDCl₃) δ: 17.7, 21.6, 24.2, 27.7, 69.0, 69.9, 83.4, 127.0,129.9, 133.0, 144.8.

Example 53

This example demonstrates the synthesis of the intermediate6-bromohex-1-yne (27) in an embodiment of the invention (FIG. 4A).

LiBr (1.7 g, 19.6 mmol) was added to a stirred solution of 26 (1.64 g,6.52 mmol) in dry DMF (20 mL). After the exothermic reaction, themixture was stirred at room temperature for 24 h. Then, water (25 mL)was added and the aqueous phase extracted with Et₂O (3×25 mL). Thecollected organic fractions were dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash chromatographyusing as eluant Hex:EtOAc (5:1) to afford a colorless oil (0.86 g, 82%).¹H NMR (400 MHz, CDCl₃) δ: 1.66-1.70 (2H, m), 1.90-1.98 (3H, m), 2.24(2H, m), 3.59 (2H, J=6.4, t). 13C NMR (100 MHz, CDCl₃) δ: 19.7, 29.4,29.7, 45.6, 69.2, 83.8.

Example 54

This example demonstrates the synthesis of the intermediate6-bromohexan-1-amine hydrochloride (29) in an embodiment of theinvention (FIG. 4A).

6-Aminohexanol 28 (0.5 g, 4.27 mmol) was slowly added to a stirring 48%hydrogen bromide solution (5.1 mL) at 0° C., and the resulting mixturewas stirred at 80° C. for 20 h. The mixture was neutralized by addingNaOH 2N (20 mL) and extracted with EtOAc (3×20 mL). The combined organicfraction was washed with water (50 mL) followed by brine (50 mL) andthen dried over Na₂SO₄, filtered, and concentrated in vacuo. Theobtained viscous oil was then dissolved in 4M hydrogen chloride solutionin dioxane to give a sticky solid that was washed with Et₂O and thenfiltered for affording to a yellowish solid (0.55 g, 61%). ¹H NMR (400MHz, MeOD) δ:1.42-1.46 (4H, m), 1.59-1.70 (2H, m), 1.74-1.82 (2H, m),2.86 (2H, J=8, t), 3.39 (2H, J=4, t). ¹³C NMR (100 MHz, MeOD) δ: 25.2,27.0, 27.2, 32.2, 32.8, 44.2.

Example 55

This example demonstrates the synthesis of the intermediate6-azidohexan-1-amine (30) in an embodiment of the invention (FIG. 4A).

To a solution of 29 (0.55 g, 2.54 mmol) in water (25 mL), NaN₃ (0.49 g,7.69 mmol) was added and the resulting mixture was heated at 100° C. for12 h. After cooling, 37% ammonia solution was added until a basic pH wasreached, and the aqueous phase was extracted with Et₂O (3×20 mL). Theorganic fractions were collected, dried over Na₂SO₄ and filtered, andthe solvent removed under vacuum to give a yellow oil (0.29 g, 80%). ¹HNMR (400 MHz, CDCl₃) δ:1.29-1.34 (4H, m), 1.35-1.40 (2H, m), 1.50-1.54(2H, m), 2.15 (2H, br s), 2.62 (2H, J=4, t), 3.18 (2H, J=8.0, t). ¹³CNMR (100 MHz, CDCl₃) δ: 26.4, 26.6, 28.8, 33.1, 41.8, 51.4

Example 56

This example demonstrates the synthesis of ethyl4-(4-(piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoate(32) in an embodiment of the invention (FIG. 4A).

To a solution of 31 (0.50 g, 1.57 mmol) in EtOH (50 mL), thionylchloride (1.37 mL, 18.84 mmol) was carefully added over 30 min at 0° C.The reaction was allowed to warm up at room temperature and stirredovernight. The resulting mixture was quenched by adding 5% solution NaOH(25 mL) until basic pH. Then, the solvent was evaporated under vacuum,and the aqueous residue was extracted with EtOAc (3×20 mL). Thecollected organic fractions were dried over Na₂SO₄, filtered and thesolvent was removed under vacuum. The residue was purified bychromatography using as eluant DCM/MeOH/NH₄OH (7:3:0.3). The titlecompound was obtained as a white solid (0.61 g, 78%). ¹H NMR (400 MHz,MeOD) δ: 1.35 (3H, J=4 Hz, t), 1.65-1.69 (2H, m), 1.81-1.84 (2H, m),2.73-2.79 (3H, m), 3.15-3.19 (2H, m), 4.35 (2H, J=8 Hz, q), 7.33-7.37(4H, m), 7.69 (2H, J=8 Hz, d), 7.76-7.79 (1H, m), 7.83-7.90 (4H, m),8.26 (1H, s), 8.58 (1H, s). m/z (ESI, MH⁺) 504; ESI-HRMS (MH⁺) calcd.for C₃₁H₂₉F₃NO₂ 504.2150, found 504.2150.

Example 57

This example demonstrates the synthesis of ethyl4-(4-(1-(hex-5-yn-1-yl)piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoate(33) in an embodiment of the invention (FIG. 4A).

K₂CO₃ (0.12 g, 0.9 mmol) was added to a stirring solution of 32 (0.15 g,0.3 mmol) in dry dimethylformamide (DMF) (15 mL), and the resultingmixture was left stirring for 20 min. Compound 26 (0.06 g, 0.6 mmol) wassubsequently added, and the reaction mixture was first stirred at roomtemperature for 2h and then at 50° C. for 2.5h. After cooling at roomtemperature, NaHCO₃ saturated solution (15 mL) was added, and theaqueous phase was extracted with EtOAc (3×20 mL). The collected organicfractions were dried over Na₂SO₄, filtered and the solvent removed undervacuum. The residue was purified by chromatography using as eluantDCM/MeOH/NH₄OH (9.5:0.5:0.05). The desired compound was obtained as acolorless oil (0.16 g, 92%). m/z ¹H NMR (400 MHz, CDCl₃) δ: 1.38 (3H,J=4 Hz, t), 1.51-1.55 (2H, m), 1.65-1.69 (2H, m), 1.90-1.99 (4H, m),2.06-2.17 (5H, m), 2.40-2.44 (2H, m), 2.56-2.59 (1H, m), 3.10-3.13 (2H,m), 4.38 (2H, J=8 Hz, q), 7.33 (2H, J=8 Hz, d), 7.38 (2H, J=8 Hz, d),7.67-7.70 (3H, m), 7.71-7.76 (2H, m), 7.91-7.95 (2H, m), 8.15 (1H, s),8.60 (1H, s) (ESI, MH⁺) 584. ¹³C NMR (101 MHz, CDCl3) δ 14.42, 18.35,25.69, 25.77, 26.49, 29.71, 32.99, 42.33, 54.27, 58.31, 61.26, 68.58,84.31, 125.89, 126.65, 126.99, 127.11, 127.44, 127.94, 128.03, 129.55,130.07, 130.61, 133.30, 137.61, 140.47, 143.91, 166.56. (ESI, MH⁺) 584;ESI-HRMS (MH⁺) calcd. for C₃₇H₃₇F₃NO₂ 584.2783, found 584.2776.

Example 58

This example demonstrates the synthesis of ethyl4-(4-(1-(4-(1-(6-aminohexyl)-1H-1,2,3-triazol-4-yl)butyl)piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoate(34) in an embodiment of the invention (FIG. 4A).

To a solution of 33 (50 mg, 0.09 mmol) in DCM:t-BuOH:H₂O (1:1:1) (2 mL),compound 30 was added, followed by copper (II) sulfate pentahydrate (15mol %) and sodium ascorbate (45 mol %). The reaction mixture was stirredfor 24 h at room temperature. The solvents were removed under vacuum andthe residue rinsed with 37% ammonia solution (5 mL) and extracted withEtOAc (3×8 mL). The collected organic fractions were dried over Na₂SO₄,filtered, and the solvent was removed under vacuum. The residue waspurified by chromatography using as eluant a gradient of DCM/MeOH/NH₄OH(from 9.5:0.5:0.05 to 7:3:0.3). The title product was obtained as awhite solid (32 mg, 51%). ¹H NMR (400 MHz, CDCl₃) δ: 1.35-1.39 (7H, m),1.58-1.65 (6H, m), 1.83-1.88 (6H, m), 2.04-2.07 (2H, m), 2.39-2.42 (3H,m), 2.65-2.71 (6H, m) 3.09-3.17 (2H, m), 4.23-4.26 (2H, m), 4.38 (2H,J=8 Hz, q), 7.21-7.23 (1H, m), 7.30-7.33 (2H, m), 7.38-7.40 (2H, m),7.66-7.70 (3H, m), 7.73-7.76 (2H, m), 7.96-7.99 (2H, m), 8.15 (1H, s),8.62 (1H, s). (ESI, MH⁺) 726; ESI-HRMS (MH⁺) calcd. for C₄₃H₅₁F₃N₅O₂726.3984, found 726.3995.

Example 59

This example demonstrates the synthesis of4-(4-(1-(4-(1-(6-aminohexyl)-1H-1,2,3-triazol-4-yl)butyl)piperidin-4-yl)phenyl)-7-(4-(trifluoromethyl)phenyl)-2-naphthoicacid (35) in an embodiment of the invention (FIG. 4A).

To a solution of 34 (32 mg, 44 μmol) in MeOH (1 mL), a solution of 0.5 MLiOH (1 mL) was added and the resulting mixture was heated at 60° C. andstirred overnight. After cooling, the solvents were evaporated and theresidue purified by preparative HPLC(R_(t)=31.08 min). The product wasobtained as a white solid by freeze drying (6.5 mg, 21%). ¹H NMR (400MHz, MeOD) δ: 1.26-1.33 (4H, m), 1.52-1.64 (6H, m), 1.81-1.86 (6H, m),2.19-2.25 (2H, m), 2.46-2.50 (2H, m), 2.66-2.70 (3H, m), 2.79 (2H, J=8Hz, t), 3.09-3.12 (2H, m), 4.30 (2H, J=8 Hz, t), 7.32 (2H, J=8 Hz, d),7.38 (2H, J=8 Hz, d), 7.66-7.70 (3H, m), 7.70-7.72 (4H, m), 7.89-7.94(4H, m), 8.27 (1H, s), 8.49 (1H, s). (ESI, MH⁺) 698; ESI-HRMS (MH⁺)calcd. for C₄₁H₄₇F₃N₅O₂ 698.3690, found 698.3682.

Example 60

This example demonstrates the synthesis of6-amino-9-(2-carboxy-4-((6-(4-(4-(4-(4-(3-carboxy-6-(4-(trifluoromethyl)phenyl)-naphthalen-1-yl)phenyl)piperidin-1-yl)butyl)-1H-1,2,3-triazol-1-yl)hexyl)carbamoyl)-phenyl)-3-iminio-5-sulfo-3H-xanthene-4-sulfonate(4) (FIG. 4B).

The coupling of the AlexaFluor488 fluorophore and pharmacophore offormula (I) was attempted by two methods, either: 1) condensation of thefluorophore as a carboxylic acid 36 to the ethyl ester of 34 followed byester saponification; or 2) by reaction of the fluorophore in situactivated as N-succinimidyl ester 37 with the carboxylic acid derivative35. In particular, to a solution of AlexaFluor 488, 35 (4.44 mg, 7.08μmol) and N,N-diisopropylethylamine (DIPEA) (1.34 μL, 7.72 μmol) in dryN,N-dimethylformamide (DMF) (400 μL), andN,N,N′,N′-tetramethyl-O—(N-succinimidyl)-uronium tetrafluoroborate(TSTU) (2.42 mg, 7.72 μmol) was added at 0° C. The resulting mixture wasallowed to warm up at room temperature and stirred for 2-3 h. Then, asolution of 34 (4.5 mg, 6.44 μmol) and DIPEA (1.30 μL, 7.08 μmol) in dryDMF (300 μL) was added and the reaction was stirred overnight at roomtemperature. After removal of the solvent, the residue was purified bypreparative HPLC (Method A, R_(t)24.9 min). The product 4 was obtainedas an orange solid after lyophilization (0.8 mg, 10%). (ESI, (M−H⁺)⁻)1212; ESI-HRMS (M−H⁺)⁻ calcd. for C₆₂H₅₇F₃N₂O₁₂S₂ 1212.3462, found1212.3459. HPLC purity 96.1% (R_(t)=5.7 min).

The second synthetic route provided compound 4 with an improved thereaction yield compared to the previous synthetic method (Kiselev etal., ACS Chem. Biol. 2014, 9, 2833-2842).

Example 61

This example demonstrates compounds of formula (I) assayed in acompetition assay by flow cytometry using Chinese hamster ovary (CHO)cells expressing P2Y₁₄R and fluorescent antagonist ligand 4 as a tracer.

The following compounds of formula (I)

in which Ar is defined in Table 1 below, were subjected to docking andmolecular dynamics simulation (10 ns) in a P2Y₁₄R homology model with 3bound, which was refined using molecular dynamics (standard protocol),as detailed below.

P2Y₁₄R models were uploaded to the “Orientations of Proteins inMembranes (OPM)” database and a suggested orientation for each structurewas provided based on the 2MeSADP-bound P2Y₁₂R orientation (PDB: 4PXZ)(Zhang et al., Nature 2014, 509, 119-122). Each receptor model was thenpositioned in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)lipid bilayer (70 A×70 A) generated by a grid-based method using the VMDMembrane Plugin tool, and overlapping lipids within 0.6 A were removedupon combining the protein membrane system (Lomize et al.,Bioinformatics 2006, 22, 623-5; and Sommer et al., Comput. Struct.Biotechnol. J. 2013, 5, e201302014). Each protein-membrane system wasthen solvated with TIP3P water using the Solvate 1.0 VMD Plugin tool andneutralized by 0.154 M Na+/Cl− counterions.

This study utilized the high-performance computational capabilities ofthe Biowulf Linux GPU cluster at the National Institutes of Health,Bethesda, Md. (http://biowulf.nih.gov). Molecular dynamics simulationswith periodic boundary conditions were carried out using NanoscaleMolecular Dynamics (NAMD) software and the CHARMM36 Force Field(Jorgensen et al., The Journal of Chemical Physics 1983, 79, 926-935;and Phillips et al., J Comput Chem 2005, 26, 1781-1802). The ligandswere parameterized by analogy using the ParamChem service (1.0.0) andimplementing the CHARMM General Force Field for organic molecules(3.0.1) (Best et al., Journal of Chemical Theory and Computation 2012,8, 3257-3273; Vanommeslaeghe et al., J Comput. Chem. 2010, 31, 671-90;Vanommeslaeghe et al., J. Chem. Inf. Model. 2012, 52, 3144-3154; andVanommeslaeghe et al., J Chem. Inf. Model. 2012, 52, 3155-3168)10,000-step conjugate gradient minimization was initially performed tominimize steric clashes. The protein and ligand atoms were kept fixedduring an initial 8 ns equilibration of the lipid and water molecules.Atom constraints were then removed and the entire system was allowed toequilibrate. The temperature was maintained at 300 K using a Langevinthermostat with a damping constant of 3 ps⁻¹. The pressure wasmaintained at 1 atm using a Berendsen barostat. An integration time stepof 1 fs was used, while hydrogen-oxygen bond lengths andhydrogen-hydrogen angles in water molecules were constrained using theSHAKE algorithm (Ryckaert et al., J Comput. Phys. 1977, 23, 327-341).VMD 1.9 was used for trajectory analysis and movie making. The PyMOLMolecular Graphics System, Version 1.6.0 Schrodinger, LLC was used formaking figures. Each structure was simulated for 30 ns withoutconstraints. Root-mean-square deviation (RMSD) plots for ligand atomsduring the 30 ns trajectories were used to compare relative ligandstability in the binding pocket during simulation, and thus used torefine initial ranking scores from ligand docking.

TABLE 1 P2Y₁₄R affinity^(a) (μM) or Compound % inhibition at IdentifierAr 3 μM 3 n/a 92.6 ± 1.2% PPTN 24a MRS4218

30.1 ± 2.1% 24b MRS4217

92.3 ± 0.5% 24c MRS4228

67.2 ± 2.2% 24d MRS4235

14.5 ± 2.1% 24e MRS4226

21.2 ± 2.9% 24f AJ139 MRS4244

26.5 ± 5.2% 24g AJ137D MRS4242

82.7 ± 1.5% 24h AJ136D MRS4241

88.8 ± 0.8% 24i

67.3 ± 2.5% 24j MRS4225

44.2 ± 1.9% 24k MRS4219

66.2 ± 2.0% 24-l AJ144 MRS4261

93.1 ± 0.8% 24m

NS 24n

NS 24o MRS4227

75.1 ± 2.8% 24p

91.5 ± 0.3% 24q

87.1 ± 0.5% 24r MRS4236

88.9 ± 4.6% 24t MRS4229

79.0 ± 2.2% 24u MRS4232

39.6 ± 7.4% 24v

85.2 ± 0.8% 24w MRS4233

13.7 ± 3.1% 24x MRS4230

86.7 ± 1.0% 24y

65.3 ± 4.5% 24z AJ138D MRS4243

49.9 ± 2.2% 24aa AJ122D MRS4237

65.2 ± 1.2% 24bb AJ134D MRS4238

37.3 ± 4.6% 24cc MRS4239

42.2 ± 4.3% 24dd

85.0 ± 0.6% 24ee AJ135D MRS4240

76.2 ± 1.7% 24ff AJ142 MRS4259

33.4 ± 1.3% 24gg AJ143 MRS4260

35.7 ± 0.7% 24hh

69.3 ± 3.8% 24ii

33.6 ± 3.5% 24jj AJ141 MRS4258

11.6 ± 1.2% 24kk AJ140 MRS4245

14.0 ± 5.1% 24-ll

13.1 ± 2.6% 24mm MRS4231

18.1 ± 5.8% 24nn AJ145 MRS4262

32.4 ± 2.7% 24oo

73.4 ± 3.1% 24pp MRS4234

6.0 ± 1.3% 24qq AJ146 MRS4263

53.1 ± 2.0% 24rr

50.1 ± 1.9% ^(a)% inhibition at 3 μM of binding of fluorescentantagonist 4 (20 nM) in P2Y₁₄R—CHO cells NS: not synthesized

The affinities of the various analogues were determined by the foregoingmethod, which provided sigmoidal concentration response curvesdisplaying a smooth concentration dependence of the inhibition. Forexample, the 5-chlorothienyl analogue 24k displayed an IC₅₀ value of0.73 μM. The rank order of potency was: 24k>24a≥24b.

Example 62

This example demonstrates the proposed synthesis of a dendron conjugatecomprising a compound of formula (I) in an embodiment of the invention.

The proposed syntheses of precursors are set forth in FIGS. 5 and 6. Thesynthesis of compound 101 was previously published in WO 93/21144;Newkome et al., J. Org. Chem. 1991, 56, 7162; and Newkome et al., J.Org. Chem. 1992, 57, 358. For compounds (e.g., 102d, 103d), the letterdefines the number of repeat units, as defined in Table 2.

TABLE 2 Compound n 102a 0 103a 106a 107a 102b 2 103b 106b 107b 102c 3103c 106c 107c 102d 4 103d 106d 107d 102e 5 103e 106e 107e 102f 6 103f106f 107f 102g 8 103g 106g 107g 102h 10 103h 106h 107h 102i 12 103i 106i107i 102j 11 103j 106j 107j 102k 14 103k 106k 107k 102l 15 103l 106l107l 102m 16 103m 106m 107m 102n 18 103n 106n 107n 102o 20 103o 106o107n 102p 22 103p 106p 107p 102q 23 103q 106q 107q 102r 24 103r 106r107r 102s 26 103s 106s 107s 102t 28 103t 106t 107t 102u 30 103u 106u107u 102v 32 103v 106v 107v 102w 34 103w 106w 107w 102x 36 103x 106x107x

Dendrons 109 and 111 comprising a lipoic acid residue as the reactivesulfur-containing moiety (X⁵) is shown in FIGS. 7 and 8. Synthesis ofdendrons 108 and 109 was previously described in WO 2005/075453; Cho etal., Chem. Mater. 2011, 23, 2665-2676; Cho et al., Langmuir 2014, 30,3883-3893; and Tsai et al., Nanoscale 2013, 5, 5390-5395.

Two proposed routes to the final dendrimer conjugates are detailed inStrategy 1 (FIGS. 9A-9C) and Strategy 2 (FIGS. 10A-10C). FIGS. 11 and 12illustrate the conjugation of a dendron conjugate of the invention to aparticle, such as a quantum dot (FIG. 11) or a gold particle (FIG. 12),through a reactive sulfur atom that is part of R⁵.

All references, including publications, patent applications, andpatents, cited herein are hereby incorporated by reference to the sameextent as if each reference were individually and specifically indicatedto be incorporated by reference and were set forth in its entiretyherein.

The use of the terms “a” and “an” and “the” and “at least one” andsimilar referents in the context of describing the invention (especiallyin the context of the following claims) are to be construed to coverboth the singular and the plural, unless otherwise indicated herein orclearly contradicted by context. The use of the term “at least one”followed by a list of one or more items (for example, “at least one of Aand B”) is to be construed to mean one item selected from the listeditems (A or B) or any combination of two or more of the listed items (Aand B), unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. All methodsdescribed herein can be performed in any suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof any and all examples, or exemplary language (e.g., “such as”)provided herein, is intended merely to better illuminate the inventionand does not pose a limitation on the scope of the invention unlessotherwise claimed. No language in the specification should be construedas indicating any non-claimed element as essential to the practice ofthe invention.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

1. A compound selected from formula (I), formula (II), formula (III),and formula (IV), wherein (a) the compound of formula (I) is of theformula:

wherein ring A is aryl, heteroaryl, or cycloalkyl; R¹ is —CO₂H,—CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate; R² is H, C₁-C₈alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl, cyanoalkyl, aryl,heteroaryl, heterocycloalkyl, —(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or—(CH₂)_(m)heterocycloalkyl; each R³ is the same or different and each isC₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl,C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈haloalkoxy, —CN, —NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴,—(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl; R⁴,R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl; and mand n are the same or different and each is 0 or an integer from 1-5; ora pharmaceutically acceptable salt thereof; (b) the compound of formula(II) is of the formula:

wherein ring A′ is aryl, heteroaryl, or cycloalkyl; R^(1′) is —CO₂H,—CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate; R^(2′) is H, C₁-C₈alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl, cyanoalkyl, aryl,heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl; each R^(3′) is the same or different andeach is C₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy,hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈haloalkyl, C₁-C₈ haloalkoxy, —CN, —NO₂, —NR^(5′)R^(6′), —C(O)R^(4′),—CO₂R^(4′), —C(O)NR⁵′R⁶, —NR^(5′)C(O)R^(4′), —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl; R^(4′), R^(5′),and R^(6′) are the same or different and each is H or C₁-C₈ alkyl; andm′ and n′ are the same or different and each is 0 or an integer from1-5; or a pharmaceutically acceptable salt thereof; (c) the compound offormula (III) is a conjugate of the formula:

or a pharmaceutically acceptable salt thereof, wherein ring A is aryl,heteroaryl, or cycloalkyl; R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or abioisostere of carboxylate; each R³ is the same or different and each isC₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl,C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈haloalkoxy, —CN, —NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴,—(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl; R⁴,R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl; X¹ isselected from the group consisting of —(CH₂)_(o)—, —C(O)—, —C(O)NH—,—OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—; X² is selected fromthe group consisting of

R⁷ is CH₂, NH, or O; X³ is a dendron; X⁴ is selected from the groupconsisting of —(CH₂)_(o)—, —C(O)—, —C(O)NH—, —OC(O)NH—, —OC(O)—,—C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X⁵ is a reactive sulfur-containing moiety; m, n, and q are the same ordifferent and each is 0 or an integer from 1-5; o is an integer from1-5; and p is 0 or an integer from 1-36; wherein X⁵ is optionally linkedto a particle; and (d) the compound of formula (IV) is a dendronconjugate of the formula:

or a pharmaceutically acceptable salt thereof, wherein ring A′ is aryl,heteroaryl, or cycloalkyl; R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or abioisostere of carboxylate; R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl,C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl,C₁-C₈ haloalkyl, cyanoalkyl, aryl, heteroaryl, heterocycloalkyl,—(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR^(5′)R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR^(5′)R⁶,—NR^(5′)C(O)R^(4′), —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl; R^(4′), R^(5′), and R^(6′) are the same ordifferent and each is H or C₁-C₈ alkyl; X^(1′) is selected from thegroup consisting of —(CH₂)_(o′)—, —C(O)—, —C(O)NH—, —OC(O)NH—, —OC(O)—,—C(O)O—, —C(S)NH—, and —SO₂—; X^(2′) is selected from the groupconsisting of

R⁷ is CH₂, NH, or O; X^(3′) is a dendron; X^(4′) is selected from thegroup consisting of —(CH₂)_(o′)—, —C(O)—, —C(O)NH—, —OC(O)NH—, —OC(O)—,—C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X^(5′) is a reactive sulfur-containing moiety; m′, n′, and q′ are thesame or different and each is 0 or an integer from 1-5; o′ is an integerfrom 1-5; and p′ is 0 or an integer from 1-36; wherein X^(5′) isoptionally linked to a particle.
 2. The compound of claim 1, wherein informula (I), ring A is phenyl, furanyl, thiazolyl, thienyl, pyrazolyl,pyridazinyl, pyridinyl, pyrazinyl, benzofuranyl, cyclopropyl, orcyclohexyl, or a pharmaceutically acceptable salt thereof.
 3. Thecompound of claim 2, wherein in formula (I), ring A is phenyl, or apharmaceutically acceptable salt thereof.
 4. The compound of claim 1,wherein in formula (I), R¹ is —CO₂H, or a pharmaceutically acceptablesalt thereof.
 5. The compound of claim 1, wherein in formula (I), R¹ isa bioisostere of carboxylate selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 6. The compound of claim1, wherein in formula (I), R² is H or C₂-C₈ alkynyl, or apharmaceutically acceptable salt thereof.
 7. The compound of claim 1,wherein in formula (I), R³ is C₁-C₈ alkyl, hydroxy, hydroxyalkyl, C₁-C₈alkoxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN, —NH₂, —CO₂R⁴, or apharmaceutically acceptable salt thereof.
 8. The compound of claim 1,wherein in formula (I), n is 0, 1, or
 2. 9. (canceled)
 10. The compoundof claim 1, wherein in formula (I), R¹ is —CO₂H; R² is H; and

is selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 11. The compound of claim1, wherein the compound is a compound of formula (II):

wherein ring A′ is aryl, heteroaryl, or cycloalkyl; R^(1′) is —CO₂H,—CO₂(C₁-C₈ alkyl), or a bioisostere of carboxylate; R^(2′) is H, C₁-C₈alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆cycloalkylalkyl, hydroxyalkyl, C₁-C₈ haloalkyl, cyanoalkyl, aryl,heteroaryl, heterocycloalkyl, —(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl; each R^(3′) is the same or different andeach is C₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy,hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈haloalkyl, C₁-C₈ haloalkoxy, —CN, —NO₂, —NR^(5′)R^(6′), —C(O)R^(4′),—CO₂R^(4′), —C(O)NR⁵′R⁶, —NR^(5′)C(O)R^(4′), —(CH₂)_(m′)aryl,—(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl; R^(4′), R^(5′),and R^(6′) are the same or different and each is H or C₁-C₈ alkyl; andm′ and n′ are the same or different and each is 0 or an integer from1-5; or a pharmaceutically acceptable salt thereof.
 12. A dendronconjugate comprising a compound of claim 1, wherein the compound offormula (I) or formula (II) is linked to a dendron through the nitrogenatom on the piperidinyl group instead of R² or R^(2′).
 13. (canceled)14. The compound of claim 1, wherein the compound is a conjugate offormula (III)

or a pharmaceutically acceptable salt thereof, wherein ring A is aryl,heteroaryl, or cycloalkyl; R¹ is —CO₂H, —CO₂(C₁-C₈ alkyl), or abioisostere of carboxylate; each R³ is the same or different and each isC₁-C₈ alkyl, C₂-C₈ alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl,C₁-C₈ alkoxy, C₃-C₆ cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈haloalkoxy, —CN, —NO₂, —NR⁵R⁶, —C(O)R⁴, —CO₂R⁴, —C(O)NR⁵R⁶, —NR⁵C(O)R⁴,—(CH₂)_(m)aryl, —(CH₂)_(m)heteroaryl, or —(CH₂)_(m)heterocycloalkyl; R⁴,R⁵, and R⁶ are the same or different and each is H or C₁-C₈ alkyl; X¹ isselected from the group consisting of —(CH₂)_(o)—, —C(O)—, —C(O)NH—,—OC(O)NH—, —OC(O)—, —C(O)O—, —C(S)NH—, and —SO₂—; X² is selected fromthe group consisting of

R⁷ is CH₂, NH, or O; X³ is a dendron; X⁴ is selected from the groupconsisting of —(CH₂)_(o)—, —C(O)—, —C(O)NH—, —OC(O)NH—, —OC(O)—,—C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X⁵ is a reactive sulfur-containing moiety; m, n, and q are the same ordifferent and each is 0 or an integer from 1-5; o is an integer from1-5; and p is 0 or an integer from 1-36; wherein X⁵ is optionally linkedto a particle.
 15. The compound of claim 14, wherein in formula (III),the dendron is carboxyethylpolyamido (CEPAM) dendron that is optionallyfunctionalized in at least one position to include the moiety

wherein R⁸ is CH₂, NH, or O, and R⁹ is —NH₂ or —CO₂H.
 16. The compoundof claim 15, wherein in formula (III), the dendron is of generation G1,G2, or G3.
 17. The compound of claim 14, wherein the compound of formula(III) is linked to a particle through a sulfur atom of X⁵.
 18. Thecompound of claim 17, wherein the particle is a quantum dot, anon-metallic particle, or a metallic particle.
 19. (canceled) 20.(canceled)
 21. The compound of claim 1, wherein the compound is adendron conjugate of formula (IV)

or a pharmaceutically acceptable salt thereof, wherein ring A′ is aryl,heteroaryl, or cycloalkyl; R^(1′) is —CO₂H, —CO₂(C₁-C₈ alkyl), or abioisostere of carboxylate; R^(2′) is H, C₁-C₈ alkyl, C₂-C₈ alkenyl,C₂-C₈ alkynyl, C₃-C₆ cycloalkyl, C₃-C₆ cycloalkylalkyl, hydroxyalkyl,C₁-C₈ haloalkyl, cyanoalkyl, aryl, heteroaryl, heterocycloalkyl,—(CH₂)_(m′)aryl, —(CH₂)_(m′)heteroaryl, or —(CH₂)_(m′)heterocycloalkyl;each R^(3′) is the same or different and each is C₁-C₈ alkyl, C₂-C₈alkenyl, C₃-C₆ cycloalkyl, hydroxy, hydroxyalkyl, C₁-C₈ alkoxy, C₃-C₆cycloalkyloxy, aryloxy, halo, C₁-C₈ haloalkyl, C₁-C₈ haloalkoxy, —CN,—NO₂, —NR^(5′)R^(6′), —C(O)R^(4′), —CO₂R^(4′), —C(O)NR⁵′R^(6′),—NR^(5′)C(O)R^(4′), —(CH₂)_(m)aryl, —(CH₂)_(m′)heteroaryl, or—(CH₂)_(m′)heterocycloalkyl; R^(4′), R^(5′), and R^(6′) are the same ordifferent and each is H or C₁-C₈ alkyl; X^(1′) is selected from thegroup consisting of —(CH₂)_(o′)—, —C(O)—, —C(O)NH—, —OC(O)NH—, —OC(O)—,—C(O)O—, —C(S)NH—, and —SO₂—; X^(2′) is selected from the groupconsisting of

R^(7′) is CH₂, NH, or O; X^(3′) is a dendron; X^(4′) is selected fromthe group consisting of —(CH₂)_(o′)—, —C(O)—, —C(O)NH—, —OC(O)NH—,—OC(O)—, —C(O)O—, —C(S)NH—, —SO₂—, —NHC(O)—, and

X^(5′) is a reactive sulfur-containing moiety; m′, n′, and q′ are thesame or different and each is 0 or an integer from 1-5; o′ is an integerfrom 1-5; and p′ is 0 or an integer from 1-36; wherein X^(5′) isoptionally linked to a particle.
 22. A pharmaceutical compositioncomprising (i) at least one compound of claim 1 or a pharmaceuticallyacceptable salt thereof and (ii) a pharmaceutically acceptable carrier.23. A method of antagonizing P2Y₁₄ receptor (P2Y₁₄R) activity in a cellcomprising contacting the cell with the compound of claim 1 or apharmaceutically acceptable salt thereof.
 24. A method of treating adisease in a subject in need thereof comprising administering to thesubject an effective amount of the compound of claim 1 or apharmaceutically acceptable salt thereof, wherein the diseases isinflammation, diabetes, insulin resistance, hyperglycemia, a lipiddisorder, obesity, a condition associated with metabolic syndrome, orasthma.